LOCUS       sequence-14179-e        5001 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 5001)
  AUTHORS   Thomas H, Jaschkowitz K, Bulman M, Frayling TM, Mitchell SM, Roosen 
            S, Lingott-Frieg A, Tack CJ, Ellard S, Ryffel GU, Hattersley AT
  TITLE     A distant upstream promoter of the HNF-4alpha gene connects the 
            transcription factors involved in maturity-onset diabetes of the 
            young.
  JOURNAL   Hum Mol Genet. 2001 Sep 15. 10(19):2089-97.
  PUBMED    11590126
REFERENCE   2  (bases 1 to 5001)
  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: "Hum Mol 
            Genet. 2001 Sep 15. 10(19):2089-97."
COMMENT     Sequence Label: HNF4A_P2-198
FEATURES             Location/Qualifiers
     source          1..5001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      265..274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             271..1923
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(337..355)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1964..2085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1975..1994)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2029..2048
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2333..2350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2504..3092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2584..2603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3263..4054,4055..4123))
                     /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     3886..3905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4124..4228)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4255..4710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4342..4361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4552..4573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4841..4889
                     /note="synthetic polyadenylation signal"
     misc_feature    4903..4994
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4943..4962
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagggt tccctaagtg actggttact
       61 ctttaacgta tccacccacc ttgggtgatt agaagaatca ataagataac cgggcggtgg
      121 cagctggccg cactcaccgc cttcctggtg gacgggctcc tggtggctgt gctgctgctg
      181 tgagcgggcc cctgctcctc catgccccca gctctccggc tgggtgggct tggccaagct
      241 tggcattccg gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc
      301 ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg
      361 aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac
      421 atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
      481 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
      541 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
      601 gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa
      661 aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt
      721 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
      781 catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag
      841 acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg
      901 cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa
      961 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     1021 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     1081 gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc
     1141 ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     1201 gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg
     1261 ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc agctattctg
     1321 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     1381 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg
     1441 tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     1501 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa
     1561 cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct
     1621 cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca
     1681 ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     1741 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     1801 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     1861 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg
     1921 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     1981 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2041 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2101 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     2161 ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa
     2221 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     2281 cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca
     2341 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2401 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2761 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2821 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2881 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2941 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3001 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3061 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3121 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3181 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3241 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3301 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3361 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3421 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3481 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3541 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3601 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3661 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3721 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3781 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3841 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3901 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3961 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4021 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4081 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4141 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4201 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     4261 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4321 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4381 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4441 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4501 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4561 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4621 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4681 attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt
     4741 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag
     4801 taagtaatat taaggtacgg gaggtacttg gagcggccgc aataaaatat ctttattttc
     4861 attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca
     4921 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
     4981 cagaacattt ctctatcgat a
//
