LOCUS       sequence-257771-        8646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PuroR and dsRed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8646)
  AUTHORS   Kandul NP, Liu J, Hsu AD, Hay BA, Akbari OS
  TITLE     A drug-inducible sex-separation technique for insects.
  JOURNAL   Nat Commun. 2020 Apr 30;11(1):2106. doi: 10.1038/s41467-020-16020-2.
  PUBMED    32355156
REFERENCE   2  (bases 1 to 8646)
  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: "Nat 
            Commun."; date: "2020-04-30"; volume: "11(1):2106. doi"; pages: " 
            10.1038/s41467-020-16020-2"
COMMENT     Sequence Label: Opie2-PuroR
FEATURES             Location/Qualifiers
     source          1..8646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..77
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    194..215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(230..235,236..253,254..260)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    268..284
                     /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     273..295
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     292..308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     292..308
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   489..523
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1212..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(1454..2128,2129..2131,2132..2134))
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1516..1539)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1714..1733)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1931..1951
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     promoter        complement(2145..2736)
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     enhancer        2740..3222
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     CDS             3785..4384
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3785..3804)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4281..4301
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4402..4536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5572..5634
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6338..6355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6338..6354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6347..6369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6563..6582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6682..6704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6742..6760)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6828..6932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6933..7001,7002..7793)
                     /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(7151..7170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7964..8552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8453..8472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
       61 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      121 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      181 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      241 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      301 gctatgacat gattacgaat tcgagctcgg tacccgggga tcctctagag tcgacgctcg
      361 cgcgacttgg tttgccattc tttagcgcgc gtcgcgtcac acagcttggc cacaatgtgg
      421 tttttgtcaa acgaagattc tatgacgtgt ttaaagttta ggtcgagtaa agcgcaaatc
      481 ttttttaacc ctagaaagat agtctgcgta aaattgacgc atgcattctt gaaatattgc
      541 tctctctttc taaatagcgc gaatccgtcg ctgtgcattt aggacatctc agtcgccgct
      601 tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt gaactataac
      661 gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt gacttttaag atttaactca
      721 tacgataatt atattgttat ttcatgttct acttacgtga taacttatta tatatatatt
      781 ttcttgttat agatatcgtg actaatatat aataaaatgg gtagttcttt agacgatgag
      841 catatcctct ctgctcttct gcaaagcgat gacgagcttg ttggtgagga ttctgacagt
      901 gaaatatcag atcacgtaag tgaagatgac gtccagagcg atacagaaga agcgtttata
      961 gatgaggtac atgaagtgca gccaacgtca agcggtagtg aaatattaga cgaacaaaat
     1021 gttattgaac aaccaggttc ttcattggct tctaacagaa tcttgacctt gccacagagg
     1081 actattagag gtaagaataa acattgttgg tcaacttcaa agtccacgag gcgtagccga
     1141 gtctctgcac tgaacattgt cagatccgag atcggccggc ctaggcgcgc cgtacgcgta
     1201 tcgataagct ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1261 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1321 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1381 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct
     1441 agagtcgcgg ccgctacagg aacaggtggt ggcggccctc ggtgcgctcg tactgctcca
     1501 cgatggtgta gtcctcgttg tgggaggtga tgtccagctt ggagtccacg tagtagtagc
     1561 cgggcagctg cacgggcttc ttggccatgt agatggactt gaactccacc aggtagtggc
     1621 cgccgtcctt cagcttcagg gccttgtgga tctcgccctt cagcacgccg tcgcgggggt
     1681 acaggcgctc ggtggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg
     1741 aggggaagtt cacgccgatg aacttcacct tgtagatgaa gcagccgtcc tgcagggagg
     1801 agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga
     1861 agccctcggg gaaggacagc ttcttgtagt cggggatgtc ggcggggtgc ttcacgtaca
     1921 ccttggagcc gtactggaac tggggggaca ggatgtccca ggcgaagggc agggggccgc
     1981 ccttggtcac cttcagcttc acggtgttgt ggccctcgta ggggcggccc tcgccctcgc
     2041 cctcgatctc gaactcgtgg ccgttcacgg tgccctccat gcgcaccttg aagcgcatga
     2101 actccttgat gacgttcttg gaggagcgca ccatctcgag gtcacttggt tgttcacgat
     2161 cttgtcgccg ccagtgtcaa cttgcaactg aaacaatatc caacatgaac gtcaatttat
     2221 actgccctaa tggcgaacac gataacaata tttcttttat tatgccctct aaaaccaacg
     2281 cggttatcgt ttatttattc aaattagata tagaacatcc gccgacatac aatgttaatg
     2341 caaaaacgcg tttggtgagc ggatacgaaa acagtcggcc gataaacatt aatctgaggt
     2401 cgataacacc gtccttgaac ggaacacgag gagcgtacgt gatcagctgc attcgcgcgc
     2461 cgcgccttta tcgagattta tttgcataca acaagtacac tgcgccgttg ggatttgtgg
     2521 taacgcgcac acatgcagag ctgcaagtgt ggcacatttt gtctgtgcgc aaaacctttg
     2581 aagccaaaag tacgaggtcc gttacgggca tgctagcgca cacggacaat ggacccgaca
     2641 aattctacgc caaggattta atgataatgt cgggcaacgt atccgttcat tttatcaata
     2701 acctacaaaa atgtcgcgcg catcacaaag acatcgacgc gcgtagaatt ctacccgtaa
     2761 agcgagttta gttatgagcc atgtgcaaaa catgacatca gcttttattt ttataacaaa
     2821 tgacatcatt tcttgattgt gttttacacg tagaattcta ctcgtaaagc gagttcagtt
     2881 ttgaaaaaca aatgacatca tctttttgat tgtgctttac aagtagaatt ctacccgtaa
     2941 atcaagttcg gttttgaaaa acaaatgagt catattgtat gatatcatat tgcaaaacaa
     3001 atgactcatc aatcgatcgt gcgttacacg tagaattcta ctcgtaaagc gagtttatga
     3061 gccgtgtgca aaacatgaca tcatctcgat ttgaaaaaca aatgacatca tccactgatc
     3121 gtgcgttaca agtagaattc tactcgtaaa gccagttcgg ttatgagccg tgtgcaaaac
     3181 atgacatcag cttatgactc atacttgatt gtgttttacg cggtttaaac gcgcccatac
     3241 tcggtggcct ccccaccacc aacttttttg cactgcaaaa aaacacgctt ttgcacgcgg
     3301 gcccatacat agtacaaact ctacgtttcg tagactattt tacataaata gtctacaccg
     3361 ttgtatacgc tccaaataca ctaccacaca ttgaaccttt ttgcagtgca aaaaagtacg
     3421 tgtcggcagt cacgtaggcc ggccttatcg ggtcgcgtcc tgtcacgtac gaatcacatt
     3481 atcggaccgg acgagtgttg tcttatcgtg acaggacgcc agcttcctgt gttgctaacc
     3541 gcagccggac gcaactcctt atcggaacag gacgcgcctc catatcagcc gcgcgttatc
     3601 tcatgcgcgt gaccggacac gaggcgcccg tcccgcttat cgcgcctata aatacagccc
     3661 gcaacgatct ggtaaacaca gttgaacagc atctgttaca gcgacacaac atgagccggt
     3721 ccaacaacgc caacgcgccc acgccatcca accgccgccg caacctgtct ctggtgttaa
     3781 ttaaatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     3841 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     3901 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     3961 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4021 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4081 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     4141 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     4201 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     4261 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     4321 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     4381 ctgagagatc ttcgacctcg aaacttgttt attgcagctt ataatggtta caaataaagc
     4441 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4501 tccaaactca tcaatgtatc ttatcatgtc tggatcatgt cgaggggggg cccgatttaa
     4561 ataaaccccg ggcgagctcg aattaaccat tgtgggaacc gtgcgatcaa acaaacgcga
     4621 gataccggaa gtactgaaaa acagtcgctc caggccagtg ggaacatcga tgttttgttt
     4681 tgacggaccc cttactctcg tctcatataa accgaagcca gctaagatgg tatacttatt
     4741 atcatcttgt gatgaggatg cttctatcaa cgaaagtacc ggtaaaccgc aaatggttat
     4801 gtattataat caaactaaag gcggagtgga cacgctagac caaatgtgtt ctgtgatgac
     4861 ctgcagtagg aagacgaata ggtggcctat ggcattattg tacggaatga taaacattgc
     4921 ctgcataaat tcttttatta tatacagcca taatgtcagt agcaagggag aaaaggttca
     4981 aagtcgcaaa aaatttatga gaaaccttta catgagcctg acgtcatcgt ttatgcgtaa
     5041 gcgtttagaa gctcctactt tgaagagata tttgcgcgat aatatctcta atattttgcc
     5101 aaatgaagtg cctggtacat cagatgacag tactgaagag ccagtaatga aaaaacgtac
     5161 ttactgtact tactgcccct ctaaaataag gcgaaaggca aatgcatcgt gcaaaaaatg
     5221 caaaaaagtt atttgtcgag agcataatat tgatatgtgc caaagttgtt tctgactgac
     5281 taataagtat aatttgtttc tattatgtat aagttaagct aattacttat tttataatac
     5341 aacatgactg tttttaaagt acaaaataag tttatttttg taaaagagag aatgtttaaa
     5401 agttttgtta ctttatagaa gaaattttga gtttttgttt ttttttaata aataaataaa
     5461 cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt
     5521 aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca
     5581 attttacgca tgattatctt taacgtacgt cacaatatga ttatctttct agggttaaat
     5641 aatagtttct aattttttta ttattcagcc tgctgtcgtg aataccgtat atctcaacgc
     5701 tgtctgtgag attgtcgtat tctagccttt ttagtttttc gctcatcgac ttgatattgt
     5761 ccgacacatt ttcgtcgatt tgcgttttga tcaaagactt gagcagagac acgttaatca
     5821 actgttcaaa ttgatccata ttaacgatat caacccgatg cgtatatggt gcgtaaaata
     5881 tattttttaa ccctcttata ctttgcactc tgcgttaata cgcgttcgtg tacagacgta
     5941 atcatgtttt cttttttgga taaaactcct actgagtttg acctcatatt agaccctcac
     6001 aagttgcaaa acgtggcatt ttttaccaat gaagaattta aagttatttt aaaaaatttc
     6061 atcacagatt taaagaagaa ccaaaaatta aattatttca acagtttaat cgaccagtta
     6121 atcaacgtgt acacagacgc gtcggcaaaa aacacgcagc ccgacgtgtt ggctaaaatt
     6181 attaaatcaa cttgtgttat agtcacggat ttgccgtcca acgtgttcct caaaaagttg
     6241 aagaccaaca agtttacgga cactattaat tatttgattt tgccccactt cattttgtgg
     6301 gatcacaatt ttgttatatt tttaaacaaa gcttggcact ggccgtcgtt ttacaacgtc
     6361 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6421 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6481 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6541 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     6601 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6661 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6721 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6781 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6841 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6901 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6961 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7021 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7081 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7141 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7201 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7261 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7321 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7381 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7441 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7501 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7561 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7621 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7681 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7741 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7801 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7861 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7921 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7981 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8041 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8101 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8161 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8221 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8281 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8341 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8401 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8461 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8521 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8581 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8641 tggata
//
