LOCUS       sequence-84090-e        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  904 bp insert from human INS gene used as a control for 
            methylation-specific PCR assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  AUTHORS   Husseiny MI, Kuroda A, Kaye AN, Nair I, Kandeel F, Ferreri K
  TITLE     Development of a quantitative methylation-specific polymerase chain 
            reaction method for monitoring beta cell death in type 1 diabetes.
  JOURNAL   PLoS One. 2012;7(10):e47942. doi: 10.1371/journal.pone.0047942. Epub
            2012 Oct 29.
  PUBMED    23144715
REFERENCE   2  (bases 1 to 4835)
  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: "2012"; volume: "7(10):e47942. doi"; pages: " 
            10.1371/journal.pone.0047942. Epub 2012 Oct 29"
COMMENT     Sequence Label: pCR2.1-INS-904
FEATURES             Location/Qualifiers
     source          1..4835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     928..999
                     /codon_start=1
                     /product="signal sequence of human preproinsulin"
                     /label=insulin signal sequence
                     /translation="MALWMRLLPLLALLALWGPDPAAA"
     primer_bind     complement(1268..1287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1269..1287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1294..1311)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1294..1310)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1303..1325)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1451..1879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1538..1557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1748..1769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2223..3017
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2277..2296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2887..2906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3035..3103,3104..3895)
                     /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     complement(3253..3272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4066..4654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4555..4574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4808..4825
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtga tggatatctg cagaattcgc
      301 cctttgtggg gacaggggtc tggggacagc agcgcaaaga gccccgccct gcagcctcca
      361 gctctcctgg tctaatgtgg aaagtggccc aggtgagggc tttgctctcc tggagacatt
      421 tgcccccagc tgtgagcagg gacaggtctg gccaccgggc ccctggttaa gactctaatg
      481 acccgctggt cctgaggaag aggtgctgac gaccaaggag atcttcccac agacccagca
      541 ccagggaaat ggtccggaaa ttgcagcctc agcccccagc catctgccga cccccccacc
      601 ccaggcccta atgggccagg cggcaggggt tgacaggtag gggagatggg ctctgagact
      661 ataaagccag cgggggccca gcagccctca gccctccagg acaggctgca tcagaagagg
      721 ccatcaagca ggtctgttcc aagggccttt gcgtcaggtg ggctcagggt tccagggtgg
      781 ctggacccca ggccccagct ctgcagcagg gaggacgtgg ctgggctcgt gaagcatgtg
      841 ggggtgagcc caggggcccc aaggcagggc acctggcctt cagcctgcct cagccctgcc
      901 tgtctcccag atcactgtcc ttctgccatg gccctgtgga tgcgcctcct gcccctgctg
      961 gcgctgctgg ccctctgggg acctgaccca gccgcagcct ttgtgaacca acacctgtgc
     1021 ggctcacacc tggtggaagc tctctaccta gtgtgcgggg aacgaggctt cttctacaca
     1081 cccaagaccc gccgggaggc agaggacctg cagggtgagc caaccgccca ttgctgcccc
     1141 tggccgcccc cagccacccc ctgctcctgg cgctcccacc cagcatgggc agaagggggc
     1201 aggaggctaa gggcgaattc cagcacactg gcggccgctc gagcatgcat ctagagggcc
     1261 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     1321 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1381 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1441 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1501 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1561 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1621 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1681 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1741 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1801 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1861 caaaaattta acgcgaattt taacaaaatt cagggcgcaa gggctgctaa aggaagcgga
     1921 acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     1981 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     2041 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     2101 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     2161 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
     2221 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2281 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2341 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2401 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2461 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2521 aggatctcct gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2581 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2641 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2701 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2761 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2821 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2881 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2941 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3001 ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3061 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3121 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3181 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3241 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3301 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3361 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3421 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3481 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3541 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3601 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3661 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3721 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3781 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3841 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3901 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4081 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4141 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4201 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4261 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4321 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4381 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4441 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4501 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4561 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4621 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4681 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4741 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4801 cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
