pAAVS1-puro-DNR

LOCUS       pAAVS1-Puro-DNR         7507 bp DNA     circular SYN 26-OCT-2024
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  AUTHORS   Ming
  TITLE     Direct Submission
  JOURNAL   Exported Oct 26, 2024 from SnapGene 6.0.2 to Vector NTI(R) format
            https://www.biofeng.com
COMMENT     VNTDATE|1072224000|
COMMENT     VNTDBDATE|1072224000|
COMMENT     VNTNAME|pAAVS1-Puro-DNR|
COMMENT     VNTAUTHORNAME|Ming|
FEATURES             Location/Qualifiers
     misc_feature    7..536
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     polyA_signal    611..1087
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1098..1114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1134..1157)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
     CDS             complement(1176..1205)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
     promoter        complement(1320..1338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1364..1567)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1568..1871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1989..2488
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2528..3127
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     misc_feature    3729..4230
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(4321..4337)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4550..5005
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5287..5391
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5392..6252
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      6423..7011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7299..7320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7335..7365
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7373..7389
                     /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)."
     primer_bind     7397..7413
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tctagacaga gcagggcctt agggaagcgg gaccctgctc tgggcggagg aatatgtccc
       61 agatagcact ggggactctt taaggaaaga aggatggaga aagagaaagg gagtagaggc
      121 ggccacgacc tggtgaacac ctaggacgca ccattctcac aaagggagtt ttccacacgg
      181 acacccccct cctcaccaca gccctgccag gacggggctg gctactggcc ttatctcaca
      241 ggtaaaactg acgcacggag gaacaatata aattggggac tagaaaggtg aagagccaaa
      301 gttagaactc aggaccaact tattctgatt ttgtttttcc aaactgcttc tcctcttggg
      361 aagtgtaagg aagctgcagc accaggatca gtgaaacgca ccagacggcc gcgtcagagc
      421 agctcaggtt ctgggagagg gtagcgcagg gtggccactg agaaccgggc aggtcacgca
      481 tccccccctt ccctcccacc ccctgccaag ctctccctcc caggatcctc tctggcaccg
      541 gttgggcatg gccaggtagc ctatgctgtg tctggacgtc ctcctgctgg tatagttatt
      601 ttaaaatcag aaggacaggg aagggagcag tggttcacgc ctgtaatccc agcaatttgg
      661 gaggccaagg tgggtagatc acctgagatt aggagttgga gaccagcctg gccaatatgg
      721 tgaaaccccg tctctaccaa aaaaacaaaa attagctgag cctggtcatg catgcctgga
      781 atcccaacaa ctcgggaggc tgaggcagga gaatcgcttg aacccaggag gcggagattg
      841 cagtgagcca agattgtgcc actgcactcc agcttggttc ccaatagacc ccgcaggccc
      901 tacaggttgt cttcccaact tgccccttgc tccataccac ccccctccac cccataatat
      961 tatagaagga cacctagtca gacaaaatga tgcaacttaa ttttattagg acaaggctgg
     1021 tgggcactgg agtggcaact tccagggcca ggagaggcac tggggagggg tcacagggat
     1081 gccacccggg atctgttcag gaaacagcta tgaccgcggc cggccgttta aaccttatcg
     1141 tcgtcatcct tgtaatccag gatatcattt gctgccagat cctcttctga gatgagtttc
     1201 tgctcgagcg gccgcgtacg cgtcggtccg ctagctagtt aagcttgaga tctggcgcgc
     1261 cggcgatcgc ggcggcagat ctcctcggta ccggatccag tcgacgaatt cccggccgcc
     1321 ctatagtgag tcgtattaca aaattctgac ggttcactaa acgagctctg cttatataga
     1381 cctcccaccg tacacgccta ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt
     1441 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     1501 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attggtgtac tgccaaaacc
     1561 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     1621 gtaaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     1681 ggggcggact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     1741 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     1801 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     1861 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     1921 attaataact agtctgaaga ggagtttacg tccagccaag cttaggatct cgacctcgaa
     1981 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     2041 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     2101 cggtaggcgc caaccgactc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     2161 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     2221 agtagcacgt ctcaccagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     2281 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
     2341 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     2401 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     2461 ctcctcttcc tcatctccgg gcctttcgac ctgcatccat ctagatctcg agcagctgaa
     2521 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     2581 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     2641 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     2701 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     2761 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     2821 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     2881 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     2941 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     3001 cttcctggag acctccgcgc cccacaacct ccccttctac gagcggctcg gcttcaccgt
     3061 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     3121 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     3181 atcgatgata tcagatcccc gggatgcaga aattgatgat ctattaaaca ataaagatgt
     3241 ccactaaaat ggaagttttt cctgtcatac tttgttaaga agggtgagaa cagagtacct
     3301 acattttgaa tggaaggatt ggagctacgg gggtgggggt ggggtgggat tagataaatg
     3361 cctgctcttt actgaaggct ctttactatt gctttatgat aatgtttcat agttggatat
     3421 cataatttaa acaagcaaaa ccaaattaag ggccagctca ttcctcccac tcatgatcta
     3481 tagatctata gatctctcgt gggatcattg tttttctctt gattcccact ttgtggttct
     3541 aagtactgtg gtttccaaat gtgtcagttt catagcctga agaacgagat cagcagcctc
     3601 tgttccacat acacttcatt ctcagtattg ttttgccaag ttctaattcc atcagaagct
     3661 ggtcgagatc cggaaccctt aatataactt cgtataatgt atgctataga agttattagg
     3721 tccactagtg ctgtcctgaa gtggacatag gggcccgggt tggaggaaga agactagctg
     3781 agctctcgga cccctggaag atgccatgac agggggctgg aagagctagc acagactaga
     3841 gaggtaaggg gggtagggga gctgcccaaa tgaaaggagt gagaggtgac ccgaatccac
     3901 aggagaacgg ggtgtccagg caaagaaagc aagaggatgg agaggtggct aaagccaggg
     3961 agacggggta ctttggggtt gtccagaaaa acggtgatga tgcaggccta caagaagggg
     4021 aggcgggacg caagggagac atccgtcgga gaaggccatc ctaagaaacg agagatggca
     4081 caggccccag aaggagaagg aaaagggaac ccagcgagtg aagacggcat ggggttgggt
     4141 gagggaggag agatgcccgg agaggaccca gacacgggga ggatccgctc agaggacatc
     4201 acgtggtgca gcgccgagaa ggaagtgctc tctagagtgt cgcatgaata acgggcagtt
     4261 atcaaatgag cacggcgaca gcctgtgaaa gtcctctcca ctgactgtag cctccaattc
     4321 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4381 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4441 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     4501 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4561 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4621 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4681 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4741 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     4801 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4861 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     4921 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4981 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5041 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5101 ctgctcccgg catccgctta cagacaagct gtgaccgtca acgggagctg catgtgtcag
     5161 aggttttcac cgtcatcacc gaaacgcgcg acccgaaagg gcctcgtgat acgcctattt
     5221 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5281 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5341 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5401 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5461 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5521 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5581 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5641 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5701 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5761 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5821 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5881 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5941 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6001 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6061 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggttc tcgcggtatc
     6121 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6181 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6241 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6301 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6361 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6541 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6601 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6661 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6721 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6781 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6841 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6901 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6961 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7081 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7141 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7201 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7261 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7321 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7381 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ccttcctctt
     7441 ccagcccttc ctctttcggg cggcagcaag atcccttctg atctaaagtc caaccggcga
     7501 gagtgat
//