LOCUS       sequence-457367-        8075 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 8075)
  AUTHORS   Fan B, Malato Y, Calvisi DF, Naqvi S, Razumilava N, Ribback S, Gores
            GJ, Dombrowski F, Evert M, Chen X, Willenbring H
  TITLE     Cholangiocarcinomas can originate from hepatocytes in mice.
  JOURNAL   J Clin Invest. 2012 Aug 1;122(8):2911-5. doi: 10.1172/JCI63212. Epub
            2012 Jul 17.
  PUBMED    22797301
REFERENCE   2  (bases 1 to 8075)
  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"; doi: "10.1172/JCI63212"; 
            journalName: "J Clin Invest"; date: "2012-08-1- 1"; volume: "122"; 
            issue: "8"; pages: "2911-5"
COMMENT     Sequence Label: pT3-EF1a-NICD1
FEATURES             Location/Qualifiers
     source          1..8075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1354..1383
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(3841..3865)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3918..3959
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3997..4014)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4003..4227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4297..4330
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     4386..4406
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(4788..4805)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4788..4804)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4797..4819)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5013..5032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5132..5154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5192..5210)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5278..5382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5383..5451,5452..6243)
                     /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(5601..5620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6414..7002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6903..6922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7156..7173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7290..7311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7326..7331,7332..7349,7350..7356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7364..7380
                     /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     7369..7391
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7388..7404
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7388..7404
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7830..7849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    7902..7935
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggcttta
     1321 aaggaaccaa ttcagtcgac tggatccacc atggagcaaa agctcatttc tgaagaggac
     1381 ttgagatcca ccatgcagca tggccagctc tggttccctg agggtttcaa agtgtcagag
     1441 gccagcaaga agaagcggag agagcccctc ggcgaggact cagtcggcct caagcccctg
     1501 aagaatgcct cagatggtgc tctgatggac gacaatcaga acgagtgggg agacgaagac
     1561 ctggagacca agaagttccg gtttgaggag ccagtagttc tccctgacct gagtgatcag
     1621 actgaccaca gacagtggac ccagcagcac ctggacgctg ctgacctgcg catgtctgcc
     1681 atggccccaa caccgcctca gggggaggtg gatgctgact gcatggatgt caatgttcga
     1741 ggaccagatg gcttcacacc cctcatgatt gcctcctgca gtggaggggg ccttgagaca
     1801 ggcaacagtg aagaagaaga agatgcacct gctgtcatct ctgacttcat ctaccagggc
     1861 gccagcttgc acaaccagac agaccgcacc ggggagaccg ccttgcactt ggctgcccga
     1921 tactctcgtt cagatgctgc aaagcgcttg ctggaggcca gtgcagatgc caacatccag
     1981 gacaacatgg gccgtactcc gttacatgca gcagtttctg cagatgctca gggtgtcttc
     2041 cagatcctgc tccggaacag ggccacagat ctggatgccc gaatgcatga tggcacaact
     2101 ccactgatcc tggctgcgcg cctggccctg gagggcatgc tggaggacct catcaactca
     2161 catgctgacg tcaatgccgt ggatgaccta ggcaagtcgg ctttgcattg ggcggccgcg
     2221 gtgaacaatg tggatgctgc tgttgtgctc ctgaagaacg gagccaacaa ggacatgcag
     2281 aacaacaagg aggagactcc cctgttcctg gccgcccgtg agggcagcta tgagactgcc
     2341 aaagtgttgc tggaccactt tgccaaccgg gacatcacgg atcacatgga ccgattgccg
     2401 cgggacatcg cacaggagcg tatgcaccac gatatcgtgc ggcttttgga tgagtacaac
     2461 ctggtgcgca gcccacagct gcatggcact gccctgggtg gcacacccac tctgtctccc
     2521 acactctgct cgcccaatgg ctacctgggc aatctcaagt ctgccacaca gggcaagaag
     2581 gcccgcaagc ccagcaccaa agggctggct tgtggtagca aggaagctaa ggacctcaag
     2641 gcacggagga agaagtccca ggatggcaag ggctgcctgt tggacagctc gagcatgctg
     2701 tcgcctgtgg actccctcga gtcaccccat ggctacttgt cagatgtggc ctcgccaccc
     2761 ctcctcccct ccccattcca gcagtctcca tccatgcctc tcagccacct gcctggtatg
     2821 cctgacactc acctgggcat cagccacttg aatgtggcag ccaagcctga gatggcagca
     2881 ctggctggag gtagccggtt ggcctttgag ccacccccgc cacgcctctc ccacctgcct
     2941 gtagcctcca gtgccagcac agtgctgagt accaatggca cgggggctat gaatttcacc
     3001 gtgggtgcac cggcaagctt gaatggccag tgtgagtggc ttccccggct ccagaatggc
     3061 atggtgccca gccagtacaa cccactacgg ccgggtgtga cgccgggcac actgagcaca
     3121 caggcagctg gcctccagca tagcatgatg gggccactac acagcagcct ctccaccaat
     3181 accttgtccc cgattattta ccagggcctg cccaacacac ggctggcaac acagcctcac
     3241 ctggtgcaga cccagcaggt gcagccacag aacttacagc tccagcctca gaacctgcag
     3301 ccaccatcac agccacacct cagtgtgagc tcggcagcca atgggcacct gggccggagc
     3361 ttcttgagtg gggagcccag tcaggcagat gtacaaccgc tgggccccag cagtctgcct
     3421 gtgcacacca ttctgcccca ggaaagccag gccctgccca catcactgcc atcctccatg
     3481 gtcccaccca tgaccactac ccagttcctg acccctcctt cccagcacag ttactcctcc
     3541 tcccctgtgg acaacacccc cagccaccag ctgcaggtgc cagagcaccc cttcctcacc
     3601 ccatcccctg agtcccctga ccagtggtcc agctcctccc cgcattccaa catctctgat
     3661 tggtccgagg gcatctccag cccgcccacc accatgccgt cccagatcac ccacattcca
     3721 gaggcattta aataaacaga gatgtgggat gcaggacccc agcttccgtt cccaagccct
     3781 gttgggagtc ctttacagtg cttcaggatg cgaattcgcg gccgcactcg agatatctag
     3841 acccagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     3901 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     3961 gtaccggtta gtaatgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     4021 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4081 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4141 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4201 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4261 gggggtatcc ccacgcgccc tgtagcggcg cattaaataa cttcgtataa tgtatgctat
     4321 acgaagttat atcgtatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4381 tccacaccct aactgacaca cattccacag aattccttga aatacatcca caggtacacc
     4441 tccaattgac tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat
     4501 tttctggaat tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga
     4561 cccactggaa ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt
     4621 ggaaaaatga cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg
     4681 tttgttaaca agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag
     4741 tgtatgtaaa cttccgactt caactgtagg gtaccgagct cgaattcact ggccgtcgtt
     4801 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4861 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4921 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4981 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5041 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5101 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5161 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5221 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5281 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5341 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5401 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5461 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5521 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5581 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5641 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5701 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5761 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5821 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5881 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5941 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6001 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6061 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6121 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6181 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6241 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6301 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6361 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6421 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6481 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6541 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6601 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6661 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6721 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6781 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6841 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6901 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6961 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7021 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7081 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7141 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7201 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7261 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7321 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7381 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac
     7441 tctagaccta tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac
     7501 tcgtttttca actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt
     7561 aggacatcta ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt
     7621 atttcactta taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag
     7681 ttgactgtgc ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc
     7741 ttctgataga ctaattgaca tcatttgagt caattggagg tgtacctgtg gatgtatttc
     7801 aaggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     7861 agcggccgct agctcgagat cttaattaaa ctagtatcga tataacttcg tataatgtat
     7921 gctatacgaa gttatatgta cgggccagat atacgcgttg acattgatta ttgactaggc
     7981 ttttgcaaaa agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     8041 aatggacctt ctaggtcttg aaaggagtgg gaatt
//
