LOCUS       sequence-14181-e        5060 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 5060)
  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 5060)
  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_P2-257
FEATURES             Location/Qualifiers
     source          1..5060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      324..333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             330..1982
                     /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(396..414)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2023..2144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2034..2053)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2088..2107
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2392..2409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2563..3151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2643..2662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3322..4113,4114..4182))
                     /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     3945..3964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4183..4287)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4314..4769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4401..4420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4611..4632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4900..4948
                     /note="synthetic polyadenylation signal"
     misc_feature    4962..5053
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5002..5021
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgaggga cctcaggtct ggccaggtcg
       61 ccattgccat ggagacagca acagtcccca gccgcgggtt ccctaagtga ctggttactc
      121 tttaacgtat ccacccacct tgggtgatta gaagaatcaa taagataacc gggcggtggc
      181 agctggccgc actcaccgcc ttcctggtgg acgggctcct ggtggctgtg ctgctgctgt
      241 gagcgggccc ctgctcctcc atgcccccag ctctccggct gggtgggctt ggccaagctt
      301 ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc
      361 cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga
      421 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca
      481 tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
      541 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
      601 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
      661 aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa
      721 aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta
      781 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
      841 atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga
      901 caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc
      961 ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa
     1021 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
     1081 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     1141 agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc
     1201 tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     1261 aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt
     1321 tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca gctattctga
     1381 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     1441 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt
     1501 gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     1561 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     1621 acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc
     1681 ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag
     1741 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     1801 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     1861 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     1921 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt
     1981 aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     2041 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2101 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2161 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     2221 tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac
     2281 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2341 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     2401 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2461 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2521 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2581 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2641 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2701 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2761 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2821 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2881 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2941 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3001 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3061 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3121 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4261 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     4321 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4381 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4441 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4501 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4561 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4621 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4681 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4741 ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg
     4801 ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt
     4861 aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca
     4921 ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa
     4981 aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc
     5041 agaacatttc tctatcgata
//
