LOCUS 3XFlag DVL2 (WT) 6906 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6906)
AUTHORS Narimatsu M, Bose R, Pye M, Zhang L, Miller B, Ching P, Sakuma R,
Luga V, Roncari L, Attisano L, Wrana JL
TITLE Regulation of planar cell polarity by Smurf ubiquitin ligases.
JOURNAL Cell. 2009 Apr 17. 137(2):295-307.
PUBMED 19379695
REFERENCE 2 (bases 1 to 6906)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2009
Apr 17. 137(2):295-307."
FEATURES Location/Qualifiers
source 1..6906
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 241..306
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
polyA_signal 2546..3168
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2745..2764)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(3192..3212)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3197..3526
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3377..3512
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3568..3587)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3569..3587)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3601..3617)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3601..3617)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3614..3636)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3625..3641
/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(3649..3679)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(3832..3849)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4003..4591)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4083..4102)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4762..5622)
/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 5385..5404
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5623..5727)
/gene="bla"
/label=AmpR promoter
rep_origin 5754..6209
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5841..5860)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6051..6072
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6335..6357
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6349..6366
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6350..6366
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 6527..6906
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattgatc tggtaccatg
241 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
301 gacaagcttc ccttcaccat ggcgggtagc agcactgggg gcggtggggt tggggagacg
361 aaggtgattt accacctgga tgaggaagag actccctacc tggtgaagat ccctgtcccc
421 gccgagcgca tcaccctcgg cgatttcaag agcgtcctgc agcggcccgc gggcgccaag
481 tactttttca agtctatgga tcaggatttc ggggtggtga aggaagaaat ttcagatgac
541 aacgcccgcc tcccctgctt caacggaagg gtggtatcct ggctggtgtc ctcagataat
601 ccccaacccg agatggcccc tccagtccat gagcctcggg cagaactggc gcctccagcc
661 ccacctttac ctcctttgcc acccgagagg accagcggca ttggggactc aaggcctcca
721 tccttccacc ctaatgtgtc cagcagccat gagaatctgg agcctgagac agaaaccgag
781 tcagtagtgt cactgaggcg ggagcggcct cgcaggagag acagcagtga gcatggcgct
841 gggggccaca ggactggtgg cccctcaagg ctggagcgcc acctggccgg atacgagagc
901 tcctctaccc tcatgaccag cgagctggag agtaccagcc tgggggactc ggacgaggag
961 gacaccatga gcaggttcag cagctccacg gagcagagca gtgcctcccg cctccttaag
1021 cgccaccggc ggcgaaggaa gcagaggcca ccccgcctgg agaggacgtc atccttcagc
1081 agcgtcacag attccacaat gtctctcaat atcatcacag tcacgctaaa catggagaag
1141 tacaacttcc tgggtatctc cattgttggc cagagcaatg agcggggaga cggaggcatc
1201 tacattggct ccatcatgaa gggtggggct gtggcggccg acgggcgcat tgagccaggg
1261 gacatgcttt tgcaggtgaa tgacatgaac tttgagaaca tgagcaacga tgacgctgtg
1321 cgggtgctga gggacattgt gcacaagcct ggccccattg tgctgactgt ggccaagtgc
1381 tgggatccct ctcctcaggc ctatttcact ctcccccgaa atgagcccat ccagccaatt
1441 gaccctgctg cctgggtgtc ccattccgcg gctctgactg gcaccttccc agcctatcca
1501 ggttcctcct ccatgagcac cattacatct ggatcgtctt tgcctgatgg ctgtgaaggc
1561 cggggtctct ccgtccatac ggacatggca tcggtgacca aggccatggc agctccagag
1621 tctggactgg aagtccggga ccgcatgtgg ctcaagatca ccatccctaa tgcctttctg
1681 ggctcggatg tggttgactg gctctaccat cacgtggagg gctttcctga gcggcgggag
1741 gcccgcaagt atgccagcgg gctgctcaaa gcaggcctga tccgacacac cgtcaacaag
1801 atcaccttct ctgagcagtg ctattacgtc ttcggagacc tcagtggtgg ctgtgagagc
1861 tacctagtca acctgtctct caatgacaac gatggctcca gtggggcttc agaccaggat
1921 accctggctc ctctgcctgg ggccaccccc tggcccctgc tgcccacttt ctcctaccaa
1981 taccctgccc cacaccccta cagcccgcag cctccaccct accatgagct ttcatcttac
2041 acctatggtg ggggcagtgc cagcagccag catagtgagg gcagccggag cagtgggtcg
2101 acacggagtg atgggggggc agggcgcacg gggaggcccg aggagcgggc ccccgagtcc
2161 aagtccggca gtggcagtga gtctgagccc tccagccgag ggggcagcct tcggcggggt
2221 ggggaagcaa gtgggactag cgatgggggc cctcctccat ccagaggctc aactgggggt
2281 gcccctaatc tccgagccca cccagggctc catccctatg gaccgccccc tggcatggcc
2341 ctcccctaca accccatgat ggtggtcatg atgcccccac ctccacctcc agtccctcca
2401 gcagtgcagc ctccgggggc ccctccagtc agagacctgg gctctgtgcc cccagaactg
2461 acagccagcc gccaaagctt ccacatggcc atgggcaatc ccagcgagtt ctttgtggat
2521 gttatgtgaa ttcaagggcg atcccgggtg gcatccctgt gacccctccc cagtgcctct
2581 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
2641 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
2701 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
2761 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
2821 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
2881 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
2941 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
3001 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
3061 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
3121 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
3181 agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
3301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
3361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
3421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
3481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcctcgatcg
3541 aggaactgaa aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat
3601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
3661 cggaagcata aagtgtaaag cctggggtgc ctaatgagcg agctaactca cattaattgc
3721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
3781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
3841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
3901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
3961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
4021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
4081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
4141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
4201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
4261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
4321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
4381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
4441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
4501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
4561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
4621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
4681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
4741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
4801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
4861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
4921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
4981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
5041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
5101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
5161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
5221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
5281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
5341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
5401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
5461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
5521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
5581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
5641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
5701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
5761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
5821 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
5881 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
5941 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
6001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
6061 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
6121 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
6181 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
6241 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
6301 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
6361 gccagtgcca agctgatcta tacattgaat caatattggc aattagccat attagtcatt
6421 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
6481 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
6541 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
6601 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
6661 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
6721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
6781 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
6841 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
6901 accatg
//
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