LOCUS       sequence-72170-e        4614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses luciferase under control of human insulin gene promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4614)
  AUTHORS   Kuroda A, Rauch TA, Todorov I, Ku HT, Al-Abdullah IH, Kandeel F, 
            Mullen Y, Pfeifer GP, Ferreri K
  TITLE     Insulin gene expression is regulated by DNA methylation.
  JOURNAL   PLoS One. 2009 Sep 9;4(9):e6953. doi: 10.1371/journal.pone.0006953.
  PUBMED    19742322
REFERENCE   2  (bases 1 to 4614)
  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: "PLoS One.";
            date: "2009-09-9"; volume: "4(9):e6953. doi"; pages: " 
            10.1371/journal.pone.0006953"
COMMENT     Sequence Label: pGL410_INS421
FEATURES             Location/Qualifiers
     source          1..4614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      466..475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             472..2124
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2168..2289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2179..2198)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2233..2252
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2537..2554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2708..3296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2788..2807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3496..4287,4288..4356))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4461..4509
                     /note="synthetic polyadenylation signal"
     misc_feature    4523..4614
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4563..4582
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctctgtg gggacagggg tctggggaca gcagcgcaaa
       61 gagccccgcc ctgcagcctc cagctctcct ggtctaatgt ggaaagtggc ccaggtgagg
      121 gctttgctct cctggagaca tttgccccca gctgtgagca gggacaggtc tggccaccgg
      181 gcccctggtt aagactctaa tgacccgctg gtcctgagga agaggtgctg acgaccaagg
      241 agatcttccc acagacccag caccagggaa atggtccgga aattgcagcc tcagccccca
      301 gccatctgcc gaccccccca ccccaggccc taatgggcca ggcggcaggg gttgacaggt
      361 aggggagatg ggctctgaga ctataaagcc agcgggggcc cagcagccct cagccctcca
      421 ggacaggctg catcagaagc ttggcaatcc ggtactgttg gtaaagccac catggaagat
      481 gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg gaccgccggc
      541 gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat cgcctttacc
      601 gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg
      661 gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag
      721 aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc
      781 ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc
      841 accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta
      901 ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc
      961 atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc
     1021 gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga
     1081 ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc
     1141 gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt
     1201 caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg
     1261 ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa
     1321 tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag
     1381 tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta
     1441 ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca
     1501 gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc
     1561 aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt
     1621 gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac
     1681 aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc
     1741 gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa
     1801 tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac
     1861 atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca
     1921 gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc
     1981 agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct
     2041 aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag
     2101 aagggcggca agatcgccgt gtaataattc tagagtcggg gcggccggcc gcttcgagca
     2161 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2221 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2281 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2341 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg
     2401 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2461 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2521 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2581 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2641 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2701 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2761 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2821 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2881 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2941 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3001 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3061 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3121 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3181 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3241 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct
     3481 aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct
     3541 atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg
     3601 cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga
     3661 tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt
     3721 gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt
     3781 gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt
     3841 cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat
     3901 attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc
     3961 cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc
     4021 cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat
     4081 acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag
     4141 tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt
     4201 gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc
     4261 ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa
     4321 gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg
     4381 aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg
     4441 ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4501 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     4561 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
