LOCUS       sequence-364417-        6596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cancer 
            Res."; date: "2009-12"; volume: "7(12)"; pages: "1937-45. Epub 2009 
            Nov 24"
COMMENT     Sequence Label: pT3-EF1a-Bmi1
FEATURES             Location/Qualifiers
     source          1..6596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        join(570..575,576..593,594..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    1146..1179
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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    4137..4170
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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             join(5223..5291,5292..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      join(6254..6596,1..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
//
