LOCUS       sequence-14186-e        7003 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 7003)
  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 7003)
  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_P2-2200
FEATURES             Location/Qualifiers
     source          1..7003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2267..2276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2273..3925
                     /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(2339..2357)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3966..4087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3977..3996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4031..4050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4335..4352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4506..5094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4586..4605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5265..6056,6057..6125))
                     /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     5888..5907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6126..6230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6257..6712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6344..6363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6554..6575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6843..6891
                     /note="synthetic polyadenylation signal"
     misc_feature    6905..6996
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6945..6964
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgaggga aggcaatgtg agacctgttg
       61 aaatagcacg ggctttgggg tcagatgatg ctgaattcca gttctgtgtt ggctccttcg
      121 tagttatgtg acctcatgac gccacttcat gcagggggtt acgtaccact taaatctcca
      181 gaacattttg ggaaattgac atagggttag caaacctccc ccacccccat ataaaaatat
      241 taatactttt tttttttctg agatggagtc tggctctgtc gccaaggctg cagtgcagta
      301 gtgcaatctt ggctcactgc aacatttgcc tcctaggttg aaacgattct cctccttcag
      361 cctcccgagt agctgagatt acaggtgctc gctgccacac ccagctaatt tttgtatttt
      421 tagtagagat gggattacac cgtgttggcc agcctggtct caaactcctg acctcaagtg
      481 atccacctgc ctcagcctcc caaactgctg ggattacagg catgagccac tacacccgac
      541 caagaatatt aatacttttt tttaaaaaat atcgaagtta tttatttatt tatttattta
      601 tttatttatt tatttatttt gagatggagt ctctctctgt tgcccaggct ggagtgcagt
      661 ggcacgatct tggctcactg caacctctgt ccccctggtt caaacgattc tcttgcctca
      721 gcctcccgtt tgccaccatg cccggctaat ttttttattt ttggtagaga cggggtttca
      781 ccatgttggt caggctggcc tcaaactcct gacctcgtga tcggtgcgcc tcggcctccc
      841 aaaatgctgg ggttacaggc gtgagccact gcacccggcc ccaaaatcta acgtttgtta
      901 tcactgttgt tttagaaaag aaaggacatt ttaatgcaac ccatgcccat ctgtaggggc
      961 tgaaggtccc tctgaggtga ttgataaagc tgaaaaaaga gactatggga agagaagagg
     1021 aatattatca attagctatt ttagaagcag ggcaaactat gagctaggta ttcctataag
     1081 gaaatgaaat aatttactaa aaaacataca gcaagaaaat aatagagctg gagttttctg
     1141 atttttctga gtccagaatt tgtttcactg ttctcatttc agctctctct aacaggatga
     1201 atgaccttga tttcagaatc cccaagcagg tggtgagatc caaaactgag acaaaagaaa
     1261 cggggctgtt ccaaaaaaaa agctaggtgg caggtgtcta acatgccagg gagctaaaac
     1321 agagtgtgtg agtttcagca gcaggttgaa tttagaatgg ggaaggagac cagaggagac
     1381 gccagacagg atgactttgt cccattggcc tggaggcagc cccatgtttc tccacccctc
     1441 atatcactca ccagtttgta atagtatctt tgaatgacga tctgattaag gtccgtctcc
     1501 tccattagtc cacaagtttc gggggtacat ctactttgct catttccata tccccagagt
     1561 ctagcacaag gcctggtaca tagtaggtgc tcaataaata tgttagatga aaggaagata
     1621 acacctctat gtactagcag tgagactcca ggcatgcaat ttctctctgt ccttcagtcc
     1681 cttcatctca aggtttaatt taaatatggt aacgcctgta tgcaactccc agcatccagt
     1741 aggcactcac taaacacagt tctccaccct ccttttttcc tctgcccctc cctcggtttt
     1801 cccactactt cctgcatggt gacacaccca tagtttggag ccataaaacc caacccaggt
     1861 tggactctca cctctccagc cccttctgct ccggccctgt cctcaaattg gggggctgat
     1921 gtccccatac acctggctct gggttcccct aaccccagag tgcaggacta ggacccgagt
     1981 ggacctcagg tctggccagg tcgccattgc catggagaca gcaacagtcc ccagccgcgg
     2041 gttccctaag tgactggtta ctctttaacg tatccaccca ccttgggtga ttagaagaat
     2101 caataagata accgggcggt ggcagctggc cgcactcacc gccttcctgg tggacgggct
     2161 cctggtggct gtgctgctgc tgtgagcggg cccctgctcc tccatgcccc cagctctccg
     2221 gctgggtggg cttggccaag cttggcattc cggtactgtt ggtaaagcca ccatggaaga
     2281 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
     2341 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
     2401 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
     2461 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
     2521 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
     2581 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
     2641 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
     2701 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
     2761 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
     2821 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
     2881 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
     2941 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
     3001 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     3061 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
     3121 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
     3181 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     3241 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     3301 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     3361 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     3421 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     3481 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     3541 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     3601 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     3661 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     3721 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     3781 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     3841 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     3901 gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc ttcgagcaga
     3961 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4021 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4081 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4141 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgac
     4201 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     4261 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4441 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4501 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4561 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4621 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4681 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4741 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4801 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4861 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4921 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4981 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5041 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5101 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5161 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5221 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5281 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5341 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5401 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5461 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5521 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5581 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5641 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5701 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5761 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5821 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5881 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5941 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6001 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6061 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6121 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6181 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6241 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6301 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6361 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6421 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6481 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6541 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6601 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6661 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
     6721 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6781 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
     6841 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     6901 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     6961 tccccagtgc aagtgcaggt gccagaacat ttctctatcg ata
//
