LOCUS       sequence-14180-e        5029 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 5029)
  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 5029)
  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: "Hum Mol 
            Genet. 2001 Sep 15. 10(19):2089-97."
COMMENT     Sequence Label: HNF4A_P2-226
FEATURES             Location/Qualifiers
     source          1..5029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      293..302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             299..1951
                     /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(365..383)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1992..2113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2003..2022)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2057..2076
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2361..2378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2532..3120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2612..2631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3291..4082,4083..4151))
                     /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     3914..3933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4152..4256)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4283..4738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4370..4389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4580..4601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4869..4917
                     /note="synthetic polyadenylation signal"
     misc_feature    4931..5022
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4971..4990
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatg gagacagcaa cagtccccag
       61 ccgcgggttc cctaagtgac tggttactct ttaacgtatc cacccacctt gggtgattag
      121 aagaatcaat aagataaccg ggcggtggca gctggccgca ctcaccgcct tcctggtgga
      181 cgggctcctg gtggctgtgc tgctgctgtg agcgggcccc tgctcctcca tgcccccagc
      241 tctccggctg ggtgggcttg gccaagcttg gcattccggt actgttggta aagccaccat
      301 ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac
      361 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
      421 ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt
      481 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
      541 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
      601 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc
      661 gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
      721 aaagctccca atcatccaaa aaattattat catggattct aaaacggatt accagggatt
      781 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt
      841 tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc
      901 tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca
      961 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
     1021 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
     1081 agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa
     1141 gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat
     1201 tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa
     1261 ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg
     1321 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     1381 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     1441 gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta
     1501 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     1561 agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct
     1621 gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca
     1681 ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc
     1741 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     1801 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     1861 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     1921 ggccaagaag ggcggaaaga tcgccgtgta attctagagt cggggcggcc ggccgcttcg
     1981 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2041 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2101 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2161 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     2221 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2281 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     2341 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2401 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2461 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2521 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2581 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2641 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2701 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2761 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2821 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2881 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2941 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3001 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3061 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3121 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3181 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3241 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3301 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3361 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3421 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3481 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3541 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3601 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3661 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3721 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3781 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3841 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3901 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3961 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4021 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4081 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4141 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4201 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4261 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4321 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4381 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4441 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4501 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4561 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4621 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4681 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc
     4741 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4801 ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga
     4861 gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     4921 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
     4981 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgata
//
