LOCUS       sequence-14183-e        5088 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5088)
  AUTHORS   Wirsing A, Johnstone KA, Harries LW, Ellard S, Ryffel GU, Stanik J, 
            Gasperikova D, Klimes I, Murphy R
  TITLE     Novel monogenic diabetes mutations in the P2 promoter of the HNF4A 
            gene are associated with impaired function in vitro.
  JOURNAL   Diabet Med. 2010 Jun . 27(6):631-5.
  PUBMED    20546279
REFERENCE   2  (bases 1 to 5088)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Diabet Med.
            2010 Jun . 27(6):631-5."
COMMENT     Sequence Label: HNF4A_P2-285/-192
FEATURES             Location/Qualifiers
     source          1..5088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      352..361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             358..2010
                     /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(424..442)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2051..2172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2062..2081)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2116..2135
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2420..2437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2591..3179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2671..2690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3350..4141,4142..4210))
                     /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     3973..3992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4211..4315)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4342..4797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4429..4448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4639..4660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4928..4976
                     /note="synthetic polyadenylation signal"
     misc_feature    4990..5081
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5030..5049
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagccc cagagtgcag gactaggacc
       61 cgagtggacc tcaggtctgg ccaggtcgcc attgccatgg agacagcaac agtccccagc
      121 cgcgggttcc gtaagtgact ggttactctt taacgtatcc acccaccttg ggtgattaga
      181 agaatcaata agataaccgg gcggtggcag ctggccgcac tcaccgcctt cctggtggac
      241 gggctcctgg tggctgtgct gctgctgtga gcgggcccct gctcctccat gcccccagct
      301 ctccggctgg gtgggcttgg ccaagcttgg cattccggta ctgttggtaa agccaccatg
      361 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
      421 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
      481 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
      541 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
      601 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
      661 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
      721 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
      781 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
      841 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
      901 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
      961 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     1021 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     1081 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     1141 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     1201 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     1261 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     1321 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     1381 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     1441 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     1501 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     1561 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     1621 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     1681 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     1741 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     1801 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     1861 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     1921 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     1981 gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga
     2041 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2101 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2161 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2221 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     2281 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2341 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     2401 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2461 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2521 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2581 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2641 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2761 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2941 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3001 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3061 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3121 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3181 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3241 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3301 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3361 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3421 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3481 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3541 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3601 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3661 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3721 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3781 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3841 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3901 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3961 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4021 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4081 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4141 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4201 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4261 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4321 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4561 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4621 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4681 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4741 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc
     4801 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4861 tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag
     4921 cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     4981 gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata
     5041 ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgata
//
