pcDNA3.1(+)/Puro/TagRFP-LL5b

LOCUS       pcDNA3.1(+)/Puro/TagRFP-LL5b                9499 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  transient/stable overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1(+)/Puro/TagRFP-LL5b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9499)
  AUTHORS   Proszynski TJ, Sanes JR
  TITLE     Amotl2 interacts with LL5beta, localizes to podosomes and regulates 
            postsynaptic differentiation in muscle.
  JOURNAL   J Cell Sci. 2013 May 15;126(Pt 10):2225-35. doi: 10.1242/jcs.121327.
            Epub 2013 Mar 22.
  PUBMED    23525008
REFERENCE   2  (bases 1 to 9499)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9499
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5111..5128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5117..5341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5387..5815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5474..5493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5684..5705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5824..5844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5829..6158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6009..6144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6071..6090
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6179..6778
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6179..6198)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6675..6695
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6938..7059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6975..6994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7029..7048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7108..7124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7108..7124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7121..7143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7132..7148
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7156..7186)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7201..7222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7339..7356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7510..8098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7590..7609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8269..9129)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8892..8911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9130..9234)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9309..9328)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatct caccatggtg agcaagggcg
      721 aggagctgat taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc
      781 accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga
      841 gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct
      901 tcatgtacgg cagcagaacc ttcatcaacc acacccaggg catccccgat ttctttaagc
      961 agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc
     1021 tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca
     1081 gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg
     1141 ccaacaccga gatgctgtac cccgctgacg gcggcctgga aggcagaacc gacatggccc
     1201 tgaagctcgt gggcgggggc cacctgatct gcaacttcaa gaccacatac agatccaaga
     1261 aacccgctaa gaacctcaag atgcccggcg tctactatgt ggaccacaga ctggaaagaa
     1321 tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact
     1381 gcgacctccc tagcaaactg gggcacaaac ttaatggcat ggacgagctg tacaagctcg
     1441 aggcagaaga tagccacatg caaaagcagc tggaattcca aaatggtagc ttagaggaag
     1501 gatttgtggt acgttccctg gagaacgagc cccaaaacat gatggagagc ctaagcccga
     1561 ggaaatactc atcaagtctg aagtttaaag ccaacgggga ctattctggc tcctatttaa
     1621 ccctctccca accggtgtct gccaagagaa gcccttctcc aatgggcacc agcgtcagga
     1681 gcagtccctc cttggccaaa atccagggaa gcaaacagtt ctgcgatgga attgataaaa
     1741 atatttccat gaaacctcct atttctttcc tcagttctgc agcctctctt ggcggctatc
     1801 ctcttgggaa agcagatttg gatcattata ctggtcgaga cagtgagagg tccaccaggc
     1861 tctcagagaa gcctccctat tccagataca gctctaggaa taaatctcat gacagtgtct
     1921 actttctagg ggggctggaa ggaagaaaaa cctctggctc gctcctgacc atgtggaacg
     1981 gaaattcctt gagctgcact ggctcgtcac ccatcagcag gtctggggca gcgagcatgc
     2041 cttcaagtcc aaagcaagtc aggaagatga accttcagga ccactcgaca cttcagccca
     2101 ggttaagcag gcacaaggag ccggcctcag aaaacgtcag tgtgagaaca aggaaatact
     2161 cagggagcag tctgagtaat atgggtgcct acagccgatc gcttcccagg ttgtataaag
     2221 ccacagacaa ccagatgtcg cctctcagcc tgcctccaag aagttccctg ggcaattcca
     2281 gaagaggtca gttaggagaa aaggatctac ctcatagcct agtggatagt gacaattacc
     2341 ttaacttttc ttctctaagc tcaggggctt caccctataa aacctgtctg tcagagggaa
     2401 atccttacgt aagctccgcc ctcagtgttc ctgccagtcc tcgtgtggct cggaagatgc
     2461 tcctggcctc cacctcctct gatgactttg atagggcttc atactcaggg accagcccaa
     2521 gtcattcctt catttctgga gagccagacc gagtgcttgt ggccaggagg aacttctctt
     2581 gtggatcaat ggagcttgat gactctgatc tggaaagcct aagacagtcg tcagaaactc
     2641 cgcagccggt ccttcgagaa aggaaaagta gcattagctc catctcggga cgcgacgacc
     2701 tgatggatta tcaccgaaga cagagggagg agagactcag ggagcaggag atggagcgac
     2761 tggagcggca gcggctggag accatcctca gtctctgtgc tgagtataca aagcctgaag
     2821 gtcgccgcct gtccgctggc accacggtgg ctgatgtgca gaaaatcaac aaggagcttg
     2881 agaagctgca gctgtcggat gaggagtctg tgttcgagga cgctctggtg tgccccgatg
     2941 ccaggtacag gtgccaccgg aagggctctc tccaggatgt tgacgttgct ggcttcggga
     3001 accttggtca cagtgccagc ttcctggccc ccaggggcag caggagcgat gaactgcttg
     3061 gtgacctcac taggacgccc ccatcatcct cagccgcgtt tctgaaagca accaatgagt
     3121 cctcctacct cagtatcctt ccgaagacct cagaggatat aggtgaggaa cagaggactc
     3181 aagagttggc tgcaatggaa gacgcccgga tggtgattct gaacaacctt gaggaacttg
     3241 agcagaaaat caaagatata aatgaccaaa tggatgaatc ttccagagag ttggatatgg
     3301 aatgcgctct tttggatgga gaacagaaat ccgagacggc tgagctcatg aaggaaaagg
     3361 agattctaga tcatctaaac cggaaaatca cggaactaga aaagaacatc gtaggcgaga
     3421 agactaagga tgccgacctg ttggatgttg agagcaaaca ctttgaagac ttggaattcc
     3481 agcagcttga gcatgagagc cgcctagatg aggaaaagga gaacttgact caacagctcc
     3541 tgcgggaggt ggcagagtat cagcggaaca tcgtggctag gaaggaaaaa atttctgcat
     3601 tgaaaaagca agccagtcac atcgttcagc aggctcagag ggaacaagat cactttgtga
     3661 aagagaagaa caatctaata atgatgctcc aaagagaaaa agagaatctt tgcaatttgg
     3721 aaaagaaata ctccagcctc actggtggga aagggttccc tattaacccc aatactctga
     3781 aagagggcta tatcagtgta aatgagatta atgagtcatg tggcaattcc acgaatctat
     3841 ccccttccac tcagttccct gctgatgctg atgctgctgt cactgagcct gccttggctg
     3901 tgccggtgag ccagccccag agttcagagc actttagaag cctggaagaa aggaaaaagc
     3961 agcacaaaga aggcctctac ctgagtgaca ctctgcctcg aaagaaaacc accccttccc
     4021 tgtccccaca cttcagcagt gccacaatgg ggaggagcac caccccgaag gctcacctcc
     4081 ctctggggca gagcaatagc tgtggcagtg tgctccctca ctcactggcg accatgacca
     4141 aagactctga gtctcggagg atgctcagag gttataatca ccaacagatg agtgaaggac
     4201 aaagacagaa acccgaattt tacagccgca cagcatctga gtcaaacgtc tacttgaata
     4261 gtttccatta cccagatcgc agctacaagg atcaggccta tgacacgttg agcctggaca
     4321 gctctgacag catggagacc agcatctctg cctgttcacc ggacaatata tcaagtgcca
     4381 gcacatccaa tattgccaga atagaagaaa tggagagact cttgaaacag gcgcacgcgg
     4441 agaagactcg gctgcttgag tccagggaac gggaaatgga agccaaaaaa cgagctctgg
     4501 aagaagaaaa acgacgccgg gaaatcctgg aaaaacgatt gcaagaagaa accagccaga
     4561 ggcagaagtt aatagaaaaa gaagtcaaaa taagagagaa acaaagggca caggctcgtc
     4621 ccctgactcg ctatctgcca gtgcggaagg aagacttcga cttgcggagt cacgtggaga
     4681 ctgcaggcca caatattgac acctgttttc atgtgtcaat cacagagaag acctgccgag
     4741 gatacctcat caaaatgggt gggaaaatta aaacgtggaa aaaacgctgg tttgtttttg
     4801 accggaataa gcgaacattt tcttactatg cagacaagca tgaggctaaa ttgaaaggcg
     4861 tgatatactt tcaagccatt gaagaagtgt attatgatca cctcaagaac gctaataaga
     4921 gccctaatcc attactcacc tttagcgtca agacacatga cagaatctat tacatggtgg
     4981 ctccttcacc agaggccatg cggatctgga tggacgttat agtgactgga gcagaaggat
     5041 atacccactt cctgttgtag gcggccgctc gagcatgcat ctagagggcc cgtttaaacc
     5101 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5161 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5221 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5281 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     5341 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     5401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     5821 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5881 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6001 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6061 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     6121 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccgggg ccgccaccat
     6181 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     6241 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     6301 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     6361 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     6421 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     6481 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6541 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6601 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6661 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6721 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt
     6781 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6841 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6901 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6961 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     7021 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7081 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7141 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7201 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7261 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7321 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7381 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7441 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7501 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7561 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7621 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7681 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7741 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7801 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7861 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7921 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7981 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8041 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8101 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8161 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8221 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8281 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8341 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8401 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8461 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8521 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8581 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8641 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8701 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8761 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8821 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8881 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8941 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9001 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9061 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9121 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9181 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9241 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     9301 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     9361 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     9421 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     9481 cgggccagat atacgcgtt
//