LOCUS Exported 4919 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCS2+ Dkk1-flag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4919) AUTHORS Semenov MV, Tamai K, Brott BK, Kuhl M, Sokol S, He X TITLE Head inducer Dickkopf-1 is a ligand for Wnt coreceptor LRP6. JOURNAL Curr Biol. 2001 Jun 26. 11(12):951-61. PUBMED 11448771 REFERENCE 2 (bases 1 to 4919) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4919 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..978 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1013..1031 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1013..1030 /label=SP6 /note="SP6 promoter, forward primer" CDS 1879..1902 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(1927..1944) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1949..2083 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1973..1992) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2027..2046 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2196..2216) /label=T3 /note="T3 promoter, forward primer" promoter complement(2196..2214) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2235..2251) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2235..2251) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2248..2270) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2259..2275 /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(2283..2313) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2328..2349 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2466..2483) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2637..3225) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2717..2736) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3396..4256) /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 4019..4038 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4257..4361) /gene="bla" /label=AmpR promoter rep_origin complement(4387..4842) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4524..4545) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4736..4755 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt 61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca 121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt 181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg 241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat 301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca 361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt 421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac 481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac 541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta 601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa 661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg 721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac 781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg 841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa 901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc 961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt 1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcacc 1081 atgatggctc tgggcgcagc gggagctacc cgggtctttg tcgcgatggt agcggcggct 1141 ctcggcggcc accctctgct gggagtgagc gccaccttga actcggttct caattccaac 1201 gctatcaaga acctgccccc accgctgggc ggcgctgcgg ggcacccagg ctctgcagtc 1261 agcgccgcgc cgggaatcct gtacccgggc gggaataagt accagaccat tgacaactac 1321 cagccgtacc cgtgcgcaga ggacgaggag tgcggcactg atgagtactg cgctagtccc 1381 acccgcggag gggacgcagg cgtgcaaatc tgtctcgcct gcaggaagcg ccgaaaacgc 1441 tgcatgcgtc acgctatgtg ctgccccggg aattactgca aaaatggaat atgtgtgtct 1501 tctgatcaaa atcatttccg aggagaaatt gaggaaacca tcactgaaag ctttggtaat 1561 gatcatagca ccttggatgg gtattccaga agaaccacct tgtcttcaaa aatgtatcac 1621 accaaaggac aagaaggttc tgtttgtctc cggtcatcag actgtgcctc aggattgtgt 1681 tgtgctagac acttctggtc caagatctgt aaacctgtcc tgaaagaagg tcaagtgtgt 1741 accaagcata ggagaaaagg ctctcatgga ctagaaatat tccagcgttg ttactgtgga 1801 gaaggtctgt cttgccggat acagaaagat caccatcaag ccagtaattc ttctaggctt 1861 cacacttgtc agagacacga ctacaaggac gacgacgaca agtaagtcga cctcgagcct 1921 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt 1981 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 2041 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 2101 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc 2161 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc 2221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 2281 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 2341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 2401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 2461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 2521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 2581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 2641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 2701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 3061 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 3421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 3481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 3541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 3601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 3661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 3721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 3781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 3841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 3901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 3961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 4021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 4081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 4141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 4201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 4261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 4321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 4381 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 4441 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 4501 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 4561 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 4621 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 4681 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 4741 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 4801 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc 4861 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacc //