LOCUS Exported 6596 bp ds-DNA circular SYN 15-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pT3-EF1a-Bmi1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6596) AUTHORS Xu CR, Lee S, Ho C, Bommi P, Huang SA, Cheung ST, Dimri GP, Chen X TITLE Bmi1 functions as an oncogene independent of Ink4A/Arf repression in hepatic carcinogenesis. JOURNAL Mol Cancer Res. 2009 Dec;7(12):1937-45. Epub 2009 Nov 24. PUBMED 19934271 REFERENCE 2 (bases 1 to 6596) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6596 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 147..166 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 400..417 /label=L4440 /note="L4440 vector, forward primer" protein_bind 534..555 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 570..600 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 608..624 /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 613..635 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 632..648 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 632..648 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1074..1093) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(1146..1179) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 1320..2498 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 1551..2489 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 2446..2466 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 2515..2534 /label=T7 /note="T7 promoter, forward primer" promoter 2515..2533 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 2612..2636 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3635..3661 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind complement(3681..3705) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3758..3799 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3837..3854) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3843..4067 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind complement(4137..4170) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 4226..4246 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" primer_bind complement(4628..4645) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4628..4644) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4637..4659) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4853..4872) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4972..4994 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5032..5050) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5118..5222 /gene="bla" /label=AmpR promoter CDS 5223..6083 /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 complement(5441..5460) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6254..246 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 61 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 301 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 361 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 421 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 481 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 541 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 601 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc 661 aagcttgcat gcctgcaggt cgactctaga cctatacagt tgaagtcgga agtttacata 721 cacttaagtt ggagtcatta aaactcgttt ttcaactact ccacaaattt cttgttaaca 781 aacaatagtt ttggcaagtc agttaggaca tctactttgt gcatgacaca agtcattttt 841 ccaacaattg tttacagaca gattatttca cttataattc actgtatcac aattccagtg 901 ggtcagaagt ttacatacac taagttgact gtgcctttaa acagcttgga aaattccaga 961 aaatgatgtc atggctttag aagcttctga tagactaatt gacatcattt gagtcaattg 1021 gaggtgtacc tgtggatgta tttcaagggg atccagacat gataagatac attgatgagt 1081 ttggacaaac cacaactaga atgcagcggc cgctagctcg agatcttaat taaactagta 1141 tcgatataac ttcgtataat gtatgctata cgaagttata tgtacgggcc agatatacgc 1201 gttgacattg attattgact aggcttttgc aaaaagcttt gcaaagatgg ataaagtttt 1261 aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg 1321 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 1381 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 1441 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 1501 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg 1561 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta 1621 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg 1681 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc 1741 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt 1801 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc 1861 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg 1921 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag 1981 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg 2041 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag 2101 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga 2161 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt 2221 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt 2281 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg 2341 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat 2401 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag 2461 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactaatac 2521 gactcactat agggagaccc aagctggcta ggtaagcttg gtaccgagct cggatccact 2581 agtccagtgt ggtggaattc tgcagatatc aacaagtttg tacaaaaaag caggctccgc 2641 ggccgccccc ttcaccatgc atcgaacaac gagaatcaag atcactgagc taaatcccca 2701 cctgatgtgt gtgctttgtg gagggtactt cattgatgcc acaaccataa tagaatgtct 2761 acattccttc tgtaaaacgt gtattgttcg ttacctggag accagcaagt attgtcctat 2821 ttgtgatgtc caagttcaca agaccagacc actactgaat ataaggtcag ataaaactct 2881 ccaagatatt gtatacaaat tagttccagg gcttttcaaa aatgaaatga agagaagaag 2941 ggatttttat gcagctcatc cttctgctga tgctgccaat ggctctaatg aagatagagg 3001 agaggttgca gatgaagata agagaattat aactgatgat gagataataa gcttatccat 3061 tgaattcttt gaccagaaca gattggatcg gaaagtaaac aaagacaaag agaaatctaa 3121 ggaggaggtg aatgataaaa gatacttacg atgcccagca gcaatgactg tgatgcactt 3181 aagaaagttt ctcagaagta aaatggacat acctaatact ttccagattg atgtcatgta 3241 tgaggaggaa cctttaaagg attattatac actaatggat attgcctaca tttatacctg 3301 gagaaggaat ggtccacttc cattgaaata cagagttcga cctacttgta aaagaatgaa 3361 gatcagtcac cagagagatg gactgacaaa tgctggagaa ctggaaagtg actctgggag 3421 tgacaaggcc aacagcccag caggaggtat tccctccacc tcttcttgtt tgcctagccc 3481 cagtactcca gtgcagtctc ctcatccaca gtttcctcac atttccagta ctatgaatgg 3541 aaccagcaac agccccagcg gtaaccacca atcttctttt gccaatagac ctcgaaaatc 3601 atcagtaaat gggtcatcag caacttcttc tggttatcca tatgacgtcc cagactatgc 3661 ctgaaagggt gggcgcgccg acccagcttt cttgtacaaa gtggttgata tccagcacag 3721 tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct 3781 cctcggtctc gattctacgc gtaccggtta gtaatgagtt taaacccgct gatcagcctc 3841 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 3901 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 3961 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 4021 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 4081 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaaataa 4141 cttcgtataa tgtatgctat acgaagttat atcgtatgca tgctttgcat acttctgcct 4201 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag aattccttga 4261 aatacatcca caggtacacc tccaattgac tcaaatgatg tcaattagtc tatcagaagc 4321 ttctaaagcc atgacatcat tttctggaat tttccaagct gtttaaaggc acagtcaact 4381 tagtgtatgt aaacttctga cccactggaa ttgtgataca gtgaattata agtgaaataa 4441 tctgtctgta aacaattgtt ggaaaaatga cttgtgtcat gcacaaagta gatgtcctaa 4501 ctgacttgcc aaaactattg tttgttaaca agaaatttgt ggagtagttg aaaaacgagt 4561 tttaatgact ccaacttaag tgtatgtaaa cttccgactt caactgtagg gtaccgagct 4621 cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac 4681 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca 4741 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt 4801 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct 4861 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 4921 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 4981 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 5041 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 5101 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 5161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 5221 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 5281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 5341 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 5401 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 5461 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 5521 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 5581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 5641 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 5701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 5761 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 5821 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 5881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 5941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 6001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 6061 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 6121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 6181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 6241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 6301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 6361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 6421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 6481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 6541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagccc //