LOCUS       sequence-14178-e        4967 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 4967)
  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 4967)
  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_P2-164
FEATURES             Location/Qualifiers
     source          1..4967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      231..240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             237..1889
                     /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(303..321)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1930..2051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1941..1960)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1995..2014
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2299..2316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2470..3058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2550..2569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3229..4020,4021..4089))
                     /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     3852..3871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4090..4194)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4221..4676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4308..4327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4518..4539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4807..4855
                     /note="synthetic polyadenylation signal"
     misc_feature    4869..4960
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4909..4928
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagcca cccaccttgg gtgattagaa
       61 gaatcaataa gataaccggg cggtggcagc tggccgcact caccgccttc ctggtggacg
      121 ggctcctggt ggctgtgctg ctgctgtgag cgggcccctg ctcctccatg cccccagctc
      181 tccggctggg tgggcttggc caagcttggc attccggtac tgttggtaaa gccaccatgg
      241 aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg
      301 ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt
      361 ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc
      421 ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca
      481 gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag
      541 ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc
      601 agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa
      661 agctcccaat catccaaaaa attattatca tggattctaa aacggattac cagggatttc
      721 agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg
      781 tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta
      841 ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg
      901 ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc
      961 cattccatca cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag
     1021 tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga
     1081 ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg
     1141 acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg
     1201 aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc
     1261 tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg
     1321 tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc
     1381 tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg
     1441 taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag
     1501 acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga
     1561 ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc
     1621 ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg
     1681 ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg
     1741 tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag
     1801 taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg
     1861 ccaagaaggg cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag
     1921 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1981 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2041 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2101 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt
     2161 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     2221 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     2281 gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4201 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4261 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4321 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4381 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4441 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4501 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4561 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4621 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca
     4681 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4741 acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc
     4801 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4861 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
     4921 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgata
//
