LOCUS       sequence-72171-e        4699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses luciferase under control of mouse insulin 2 gene promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4699)
  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 4699)
  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_Ins2500
FEATURES             Location/Qualifiers
     source          1..4699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      551..560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             557..2209
                     /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    2253..2374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2264..2283)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2318..2337
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2622..2639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2793..3381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2873..2892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3581..4372,4373..4441))
                     /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    4546..4594
                     /note="synthetic polyadenylation signal"
     misc_feature    4608..4699
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4648..4667
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcggac cattaagtgc cttgctgcct gaattctgct
       61 ttccttctac ctctgagaga gagctgggga ctcggctgag ttaagaaccc agctatcaat
      121 tggaactgtg aaacagtcca agggacaaag atactaggtc cccaactgca acttcctggg
      181 gaatgatgtg gaaaaatgct cagccaagga caaagaaagc atcacccact ctggaacaat
      241 gtcccctgct gtgaactggt tcatcaggcc atcagggccc cttgttaaga ctctaattac
      301 cctaggacta agtagaggtg ttgacgtcca atgagcgctt tctgcagacc tagcaccagg
      361 gaagtgtttg gaaactgcag cttcagcccc tctggccatc tgctgaccta ccccacctgg
      421 agcccttaat gggtcaaaca gcaaagtcca gggggcagag aggaggtgct ttggtctata
      481 aaggtagtgg ggacccagta accaccagcc ctaagtgatc caagcttggc aatccggtac
      541 tgttggtaaa gccaccatgg aagatgccaa aaacattaag aagggcccag cgccattcta
      601 cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct
      661 ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga
      721 gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa
      781 ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc
      841 cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct
      901 gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa
      961 gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa
     1021 gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg
     1081 cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat
     1141 catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc
     1201 ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac
     1261 cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta
     1321 cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg
     1381 cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt
     1441 cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg
     1501 cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg
     1561 catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg
     1621 ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga
     1681 cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc
     1741 catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga
     1801 cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt
     1861 ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga
     1921 gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga
     1981 cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga
     2041 gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg
     2101 tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat
     2161 ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat aattctagag
     2221 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2281 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2341 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2401 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2461 tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     2521 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     2581 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     2641 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2701 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3541 tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg
     3601 aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga ctccccgtcg
     3661 tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca atgatgccgc
     3721 gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca gggagggccg
     3781 agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc tgctgtcgtg
     3841 atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc attgctactg
     3901 gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt tcccagcggt
     3961 caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc ttagggcctc
     4021 cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg gcagcactac
     4081 acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa
     4141 ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg gcgtctatac
     4201 gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg aatcgttctt
     4261 cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata tagcccactc
     4321 ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa
     4381 caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt tggatgctca
     4441 tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac gtctctcaag
     4501 gataagtaag taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta
     4561 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
     4621 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
     4681 ggtgccagaa catttctct
//
