LOCUS       sequence-198993-        9535 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 9535)
  AUTHORS   Ritz-Laser B, Gauthier BR, Estreicher A, Mamin A, Brun T, Ris F, 
            Salmon P, Halban PA, Trono D, Philippe J
  TITLE     Ectopic expression of the beta-cell specific transcription factor 
            Pdx1 inhibits glucagon gene transcription.
  JOURNAL   Diabetologia. 2003 Jun . 46(6):810-21.
  PUBMED    12783165
REFERENCE   2  (bases 1 to 9535)
  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: 
            "Diabetologia. 2003 Jun . 46(6):810-21."
COMMENT     Sequence Label: pWPT-PDX1
FEATURES             Location/Qualifiers
     source          1..9535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(861..1072)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    1129..1246
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1513..1557)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1742..1975
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2472..2597
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2644..3277
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    3445..3579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3469..3488)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3523..3542
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4004..4024)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4154..4219
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        complement(5353..5682)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5367..5502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5667..5687
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(5757..5774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5928..6516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6008..6027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6687..7478,7479..7547))
                     /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     7310..7329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7548..7652)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7720..7738
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7776..7798)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7898..7917
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8131..8150
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        8440..8458
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     8440..8457
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     LTR             8483..8663
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     protein_bind    8683..8716
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    8860..9448
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8966..8977
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     9375..9395
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     9461..9477
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     9462..9478
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgctggaggg ctgtggcgac gcgctaaggc caggcggcag gcggccctct cgggcggcaa
       61 cgggggcagc gggcggcaca gcacctccgg ggggcggcgg cggcggcagc gccagaagct
      121 cctcgccgga ggtcacggcg cagtcctgct caggctccgc gaccccgcca cccccgacag
      181 ctgtcccgcc gccgcgcttc ttgtcctcct cctttttcca cttcatgcgg cggttttgga
      241 accagatctt gatgtgtctc tcggtcaagt tcaacatgac agccagctcc acccggcgcg
      301 gccgtgagat gtacttgttg aataggaact ccttctccag ctctagcagc tgtgcgcgcg
      361 tgtaggccgt gcgcgtccgc ttgttctcct ccggctccgc agcgtaggcg ccgcctgccc
      421 actggccttt ccacgcgtga gctttggtag acttcatcca tgggaaaggc agctggacgc
      481 ggttgggctc ctccaggacg cccggctcgg ctccctccgg gaagggcccg gcgggcgggt
      541 gggggagcgc gagctgagcc gggaggtggt ggtgaaggtg cgccaccgcg gggtcgtcgg
      601 cgaggggggg cacctcgtac ggggagatgt ccggggggct gccctgctcc agcgcgccca
      661 gggcgccagg gaacgggtgc ggcggcggcg gcgggggctg gcggcccatg tacaggcacg
      721 caggggggct ggcgctgaac tccggcgccg ggcctcgctg gaacgcgcat gggtccttgt
      781 aaagctgcgt ggccgcgtag tactgctcct cgccgttcat ggccgaattc ctgcagcccg
      841 ggggatccgc gtcacgacac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc
      901 acggcgacta ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag
      961 tacacgacat cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg
     1021 ccgacccctc cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg
     1081 gcaccggagc catcgatcaa gcttctcgag gctagtctcg tgatcgataa aattttgaat
     1141 ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat
     1201 tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaagtta accgataccg
     1261 tcgagatccg ttcactaatc gaatggatct gtctctgtct ctctctccac cttcttcttc
     1321 tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     1381 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     1441 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     1501 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     1561 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     1621 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     1681 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     1741 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     1801 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     1861 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     1921 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     1981 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     2041 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     2101 tcctcctcca ggtctgaaga tcagcggccg gccgcttgct gtgcggtggt cttacttttg
     2161 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     2221 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     2281 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     2341 ccaggattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     2401 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     2461 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     2521 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     2581 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     2641 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     2701 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     2761 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     2821 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     2881 gtcccgccca ggccacgcct ccctggaaag tccccagcgg aaagtccctt gtagcaagct
     2941 cgatatcagc agttcttgaa gtactccgga tgcagctctc gggccacgtg atgaaatgct
     3001 aggcggctgt caaacctcca ctctaacact tctctctccg ggtcatccat cccatgcagg
     3061 ctcacagggt gtaacaagct ggtgttctct cctttattgg cctcttctac cttatctggc
     3121 tcaactggta ctagcttgta gcaccatcca aaggtcagtg gatatctgac ccctggccct
     3181 ggtgtgtagt tctgctaatc agggaagtag ccttgtgtgt ggtagatcca cagatcaagg
     3241 atatcttgtc ttctttggga gtgaattagc ccttccaact actaagtttg tagtacatat
     3301 ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg tttagagtaa
     3361 atgaaaccac aggtaattgg cagtggtaat agggtatggg gtgggaagtt tgggatgatt
     3421 ttggttagct tgagttatcc agttgatcca gacatgataa gatacattga tgagtttgga
     3481 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3541 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3601 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3661 aaatgtggta tggctgatta tgatctctag tcaaggcact atacatcaaa tattccttat
     3721 taaccccttt acaaattaaa aagctaaagg tacacaattt ttgagcatag ttattaatag
     3781 cagacactct atgcctgtgt ggagtaagaa aaaacagtat gttatgatta taactgttat
     3841 gcctacttat aaaggttaca gaatattttt ccataatttt cttgtatagc agtgcagctt
     3901 tttcctttgt ggtgtaaata gcaaagcaag caagagttct attactaaac acagcatgac
     3961 tcaaaaaact tagcaattct gaaggaaagt ccttggggtc ttctaccttt ctcttctttt
     4021 ttggaggagt agaatgttga gagtcagcag tagcctcatc atcactagat ggcatttctt
     4081 ctgagcaaaa caggttttcc tcattaaagg cattccacca ctgctcccat tcatcagttc
     4141 cataggttgg aatctaaaat acacaaacaa ttagaatcag tagtttaaca cattatacac
     4201 ttaaaaattt tatatttacc ttagagcttt aaatctctgt aggtagtttg tccaattatg
     4261 tcacaccaca gaagtaaggt tccttcacaa agatccgggg cccactcata aatccagttg
     4321 ccgccacggt agccaatcac cgtatcgtat aaatcatcgt cggtacgttc ggcatcgctc
     4381 atcacaatac gtgcctggac gtcgaggatt tcgcgtgggt caatgccgcg ccagatccac
     4441 atcagacggt taatcatgcg ataccagtga gggatggttt taccatcaag ggccgactgc
     4501 acaggcggtt gtgcgccgtg attaaagcgg cggactagcg tcgaggtttc aggatgttta
     4561 aagcggggtt tgaacagggt ttcgctcagg tttgcctgtg tcatggatgc agcctccaga
     4621 atacttactg gaaactattg taacccgcct gaagttaaaa agaacaacgc ccggcagtgc
     4681 caggcgttga aaagattagc gaccggagat tggcgggacg aatacgacgc ccatatccca
     4741 cggctgttca atccaggtat cttgcgggat atcaacaaca tagtcatcaa ccagcggacg
     4801 accagccggt tttgcgaaga tggtgacaaa gtgcgctttt ggatacattt cacgaatcgc
     4861 aaccgcagta ccaccggtat ccaccaggtc atcaataacg atgaagcctt cgccatcgcc
     4921 ttctgcgcgt ttcagcactt taagctcgcg ctggttgtcg tgatcgtagc tggaaataca
     4981 aacggtatcg acatgacgaa tacccagttc acgcgccagt aacgcacccg gtacagaccg
     5041 ccacggctta cggcaataat gcctttccat tgttcagaag gcatcagtcg gcttgcgagt
     5101 ttacgtgcat ggatctgcaa catgtcccag gtgacgatgt atttttcgct catgtgaagt
     5161 gtcccagcct gtttatctac ggcttaaaaa gtgttcgagg ggaaaatagg ttgcgcgaga
     5221 ttatagagat ctggcgcact aaaaaccagt atttcacatg agtccgcgtc tttttacgca
     5281 ctgcctctcc ctgacgcggg ataaagtggt attctcaaac atatctcgca agcctgtctt
     5341 gtgtccaagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     5401 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5461 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5521 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5581 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5641 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctgcattaa
     5701 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6361 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6961 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7621 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7681 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7741 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7801 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7861 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     7921 agagtgcacc attcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     7981 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     8041 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     8101 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     8161 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctggctagc
     8221 gatgaccctg ctgattggtt cgctgaccat ttccgggtgc gggacggcgt taccagaaac
     8281 tcagaaggtt cgtccaacca aaccgactct gacggcagtt tacgagagag atgatagggt
     8341 ctgcttcagt aagccagatg ctacacaatt aggcttgtac atattgtcgt tagaacgcgg
     8401 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaat
     8461 acacggaatt aattctagat gctgctagag attttccaca ctgactaaaa gggtctgagg
     8521 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     8581 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     8641 atctggtcta accagagaga cccagtacaa gcaaaaagca gcataacttc gtataatgta
     8701 tgctatacga agttattccc atggtgaatt gatcccatct tgtcttcgtt gggagtgaat
     8761 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     8821 agtcattggt cttaaaggta ccgagctcga attcccgatg cggggaggcg gcccaaaggg
     8881 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     8941 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     9001 cgcgcagaat ccaggtggca acacaggcga gcagccatgg aaaggacgtc agcttccccg
     9061 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     9121 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     9181 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     9241 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     9301 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     9361 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     9421 cacaaatttt gtaatccaga ggttgattat cggaattccc tcgaggtcga cggtatcgat
     9481 aagcttgaat ggacagtgtg atcctatcgt ggttcctgcg gccgccgagg cgcga
//
