LOCUS Exported 7969 bp ds-DNA circular SYN 15-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Stat3-C Flag pRc/CMV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7969) AUTHORS Bromberg JF, Wrzeszczynska MH, Devgan G, Zhao Y, Pestell RG, Albanese C, Darnell JE Jr TITLE Stat3 as an oncogene. JOURNAL Cell 1999 Aug 6;98(3):295-303. PUBMED 10458605 REFERENCE 2 (bases 1 to 7969) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7969 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 296..315 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(534..638) /gene="bla" /label=AmpR promoter primer_bind complement(713..732) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 904..1283 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1284..1487 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1438..1458 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 1532..1551 /label=T7 /note="T7 promoter, forward primer" promoter 1532..1550 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 1641..1657 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 1641..1657 /label=pBluescriptSK /note="For pBluescript vector" CDS 4065..4088 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(4097..4115) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4098..4115) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4135..4152) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4141..4365 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4411..4924 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4498..4517) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4708..4729 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4947..4967) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4952..5281 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5132..5267 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5194..5213 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5348..6142 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5402..5421) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6012..6031 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6316..6450 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6353..6372) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6407..6426 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6481..6497) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6481..6497) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6494..6516) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6505..6521 /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(6529..6559) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6574..6595 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6712..6729) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6883..7471) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6963..6982) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7642..533) /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" ORIGIN 1 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 61 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 661 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 721 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 781 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 841 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 901 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 961 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 1021 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 1081 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 1141 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 1201 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 1261 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 1321 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 1381 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 1441 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 1501 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 1561 cttggtaccg agctcggatc cactagtaac ggccgccagt gtgctggaat tctgcagata 1621 tccatcacac tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattccgcc 1681 gcgaccagcg caggccggcc agtcgggcct cagccccgga gacagtcgag acccctgact 1741 gcagcaggat ggctcagtgg aaccagctgc agcagctgga cacacgctac ctgaagcagc 1801 tgcaccagct gtacagcgac agcttcccca tggagctgcg gcagttcctg gcaccttgga 1861 ttgagagtca agactgggca tatgcagcca gcaaagagtc acatgccacg ttggtgtttc 1921 ataatctctt gggtgaaatt gaccagcaat atagccgatt cctgcaagag tccaatgtcc 1981 tctatcagca caaccttcga agaatcaagc agtttctgca gagcaggtat cttgagaagc 2041 caatggaaat tgcccggatc gtggcccgat gcctgtggga agagtctcgc ctcctccaga 2101 cggcagccac ggcagcccag caagggggcc aggccaacca cccaacagcc gccgtagtga 2161 cagagaagca gcagatgttg gagcagcatc ttcaggatgt ccggaagcga gtgcaggatc 2221 tagaacagaa aatgaaggtg gtggagaacc tccaggacga ctttgatttc aactacaaaa 2281 ccctcaagag ccaaggagac atgcaggatc tgaatggaaa caaccagtct gtgaccagac 2341 agaagatgca gcagctggaa cagatgctca cagccctgga ccagatgcgg agaagcattg 2401 tgagtgagct ggcggggctc ttgtcagcaa tggagtacgt gcagaagaca ctgactgatg 2461 aagagctggc tgactggaag aggcggcagc agatcgcgtg catcggaggc cctcccaaca 2521 tctgcctgga ccgtctggaa aactggataa cttcattagc agaatctcaa cttcagaccc 2581 gccaacaaat taagaaactg gaggagctgc agcagaaagt gtcctacaag ggcgacccta 2641 tcgtgcagca ccggcccatg ctggaggaga ggatcgtgga gctgttcaga aacttaatga 2701 agagtgcctt cgtggtggag cggcagccct gcatgcccat gcacccggac cggcccttag 2761 tcatcaagac tggtgtccag tttaccacga aagtcaggtt gctggtcaaa tttcctgagt 2821 tgaattatca gcttaaaatt aaagtgtgca ttgataaaga ctctggggat gttgctgccc 2881 tcagagggtc tcggaaattt aacattctgg gcacgaacac aaaagtgatg aacatggagg 2941 agtctaacaa cggcagcctg tctgcagagt tcaagcacct gacccttagg gagcagagat 3001 gtgggaatgg aggccgtgcc aattgtgatg cctccttgat cgtgactgag gagctgcacc 3061 tgatcacctt cgagactgag gtgtaccacc aaggcctcaa gattgaccta gagacccact 3121 ccttgccagt tgtggtgatc tccaacatct gtcagatgcc aaatgcttgg gcatcaatcc 3181 tgtggtataa catgctgacc aataacccca agaacgtgaa cttcttcact aagccgccaa 3241 ttggaacctg ggaccaagtg gccgaggtgc tcagctggca gttctcgtcc accaccaagc 3301 gagggctgag catcgagcag ctgacaacgc tggctgagaa gctcctaggg cctggtgtga 3361 actactcagg gtgtcagatc acatgggcta aattctgcaa agaaaacatg gctggcaagg 3421 gcttctcctt ctgggtctgg ctagacaata tcatcgacct tgtgaaaaag tatatcttgg 3481 ccctttggaa tgaagggtac atcatgggtt tcatcagcaa ggagcgggag cgggccatcc 3541 taagcacaaa gcccccgggc accttcctac tgcgcttcag cgagagcagc aaagaaggag 3601 gggtcacttt cacttgggtg gaaaaggaca tcagtggcaa gacccagatc cagtctgtag 3661 agccatacac caagcagcag ctgaacaaca tgtcatttgc tgaaatcatc atgggctata 3721 agatcatgga ttgtacctgc atcctggtgt ctccacttgt ctacctctac cccgacattc 3781 ccaaggagga ggcatttgga aagtactgta ggcccgagag ccaggagcac cccgaagccg 3841 acccaggtag tgctgccccg tacctgaaga ccaagttcat ctgtgtgaca ccaacgacct 3901 gcagcaatac cattgacctg ccgatgtccc cccgcacttt agattcattg atgcagtttg 3961 gaaataacgg tgaaggtgct gagccctcag caggagggca gtttgagtcg ctcacgtttg 4021 acatggatct gacctcggag tgtgctacct cccccatgtc tagagactac aaggacgacg 4081 atgacaaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga 4141 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 4201 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 4261 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 4321 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 4381 gaaccagctg gggctcgagg ggggatcccc acgcgccctg tagcggcgca ttaagcgcgg 4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 4621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 4741 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 4801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta 4861 caatttaaat atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg 4921 gggtgggtac cgagctcgaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 4981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 5041 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 5101 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 5161 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 5221 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 5281 aaagctcccg ggagcttgga tatccatttt cggatctgat caagagacag gatgaggatc 5341 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 5401 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 5461 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 5521 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 5581 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 5641 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 5701 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 5761 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 5821 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 5881 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 5941 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 6001 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 6061 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 6121 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg 6181 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 6241 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 6301 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 6361 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 6421 ctcatcaatg tatcttatca tgtctggatc cgtcgacctc gagagcttgg cgtaatcatg 6481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtg //