LOCUS       sequence-433264-        9192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HDR plasmid for inserting Ceratitis capitata transformer female 
            intron into the white pupae gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9192)
  TITLE     Engineered Genetic Sexing Strain for the Medfly via CRISPR Mediated 
            Translocation of an Alternately Spliced Intron into a Dominant 
            Visible Gene.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9192)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 1167A
FEATURES             Location/Qualifiers
     source          1..9192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   624..686
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1004..1389
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     protein_bind    4548..4933
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     CDS             join(5481..5483,5484..5486,5487..6161)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(5664..5684)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5882..5901
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6076..6099
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    6275..6396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6312..6331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6366..6385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    6424..6493
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   6963..6997
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(7004..7022)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7043..7059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7043..7059)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7056..7078)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7067..7083
                     /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)."
     promoter        complement(join(7091..7097,7098..7115,7116..7121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7136..7157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7274..7291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7445..8033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7525..7544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8204..8995,8996..9064))
                     /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     8827..8846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9065..9169)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtagat taaccctaga aagataatca tattgtgacg tacgttaaag
      661 ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat
      721 taatttgaat agatattaag ttttattata tttacactta catactaata ataaattcaa
      781 caaacaattt atttatgttt atttatttat taaaaaaaaa caaaactttc gttgatagaa
      841 gcatcctcat cacaagatga taataagtat accatcttag ctggcttcgg tttatatgag
      901 acgagagtaa ggggtccgtc aaaacaaaac atcgatgttc ccactggcct ggagcgactg
      961 tttttcagta cttccggtat ctcgcgtttg tttgcctgca ggtcacgtaa taagtgtgcg
     1021 ttgaatttat tcgcaaaaac attgcatatt ttcggcaaag taaaattttg ttgcatacct
     1081 tatcaaaaaa taagtgctgc atacttttta gagaaaccaa ataatttttt attgcatacc
     1141 cgtttttaat aaaatacatt gcataccctc ttttaataaa aaatattgca tactttgacg
     1201 aaacaaattt tcgttgcata cccaataaaa gattattata ttgcataccc gtttttaata
     1261 aaatacattg cataccctct tttaataaaa aatattgcat acgttgacga aacaaatttt
     1321 cgttgcatac ccaataaaag attattatat tgcatacctt ttcttgccat accatttagc
     1381 cgatcaattg gcgcgcctcc gaccgtccat cgcaataaaa tgagccatta tgagatcgaa
     1441 agggtctacg aaaatccgtt cggaaaaaat cagaaaatca tcaaagccga atataattaa
     1501 aatgtattac tagctaaaga aatcatcact aatatagaat gtagaatgaa cccatgtata
     1561 ttagatacta attgtatcgt aagactttca aaagtctaca agacattaaa tgacaagttg
     1621 actttaaatt tcaaataaat aatttatttt ttctataagc aataacattt ttgctaaatt
     1681 aagacttggt aattaggtaa tactattgtt gttctatgga atattcgatc gaaacattct
     1741 tatcagtctc aaaaacttaa aacaaactta taatataacc catatgcacg acacgaattg
     1801 ccaaagcaat gttcgctggt aaaagaagaa gataggtagg catattgtaa tttaaataaa
     1861 tatttcacct ttagatttga aaaatttctt tatttgtact tacagttgtc ttgctttgac
     1921 atataatgaa actgcatact gtgagtggaa ttacaatggt ctgggtattg attatcaaat
     1981 acttgctggt cccaccttca tattggtttt cacaattgct ggtgttttta tgggtttcgc
     2041 tgcagataaa tataatcgcg ttaaaatgtt aacagtttgt acattgattt ttgcagtagc
     2101 cattatatta caaggtaccg tagaagcgta ctggcagctg ttgctgttgc gtatggtcat
     2161 ggcgttgggg taagcgcttt tctcttaata ataagaaata acaacagatt tattaatttc
     2221 tctacttccc caacagtgaa tccggttgca acccactggc tacaggcatt atgtccgaca
     2281 tttttcccga aaataagcgc gcccttgtta tggccatttt caattggggc atctatggtg
     2341 gttatggcat tgcttttccc gtgggtcgtt atattaccaa atcgaatttc ttcaaccttg
     2401 gctggcgtat gtgctatttg ggcactggtg ttttggctgt attgctggcc gcactcacag
     2461 gcaccacact caaggaacca gaacgtaaag cgattggtaa ttttaaaagc atattttttt
     2521 ctttgaaatt cataagttat caattatcga tggaaatgta ttctatggag aacgttttac
     2581 ccgatgaatg ggtgcaaaaa ttattttacc ttcaaatcta caatcaacac acgctaactt
     2641 ttgtgacttg atcaactctc acctggaaaa gcaaccaact acaatcaaca ttctatggga
     2701 taatcgacaa gtgagtaaaa ttatagccgg acctcttagt acagtgtatt taaaagggga
     2761 ataatattct atcaatagga ataaaaataa ggtcagcagc catgactttt ccatcatttt
     2821 gaatatacct tatttgtttc gggattaatt gggggtcgga aatcctcttg aattcagaaa
     2881 cgggaaccgg aggaaggtgc cggtctttca gaaagctgtg aaaaatacca acatttctgc
     2941 tgccaagagc tcaataagaa gtttcaaaaa ttgtcttgga tgttgcagct gtggctgcta
     3001 agtaataaga catctattag tatctagatt tgttagacca tttaacatag tgttttaaac
     3061 gatggggtta atagatgagg gttaagaagc tagttatatt actgttgctg taacgccttc
     3121 aattgtcggt tacagagcaa acattattga atgttaatgt aaagagttta tttgttttct
     3181 agtaaacata tagcgattgg ttagtaatca ctaatagaaa tttttcataa gtatcaaaaa
     3241 agtaaacctc tttttcagtc tatgtaataa gtaaaccaag gaaagggaaa atatctacaa
     3301 tcaacaagcc attgttgcag caacaaagca actgaaacta caatcaacat tcaataaact
     3361 tgggtaattt ggaatttaat tctctgggac acctgtggat tacaacaatc aactcgaaac
     3421 ttattataca atgtaaataa aaattgatat gcatacatga agatcaagtg aaattccatt
     3481 tagaatcaat ttttttcgaa tattaagttt cttgctttaa tttatctgaa agtaaataga
     3541 cattccaaat tcaagttaac aaattaataa tgaattgact agtgattttt aagagaaaaa
     3601 gataagattt aaaaaaggaa agcctttctt gataaatttt tgaaccactt tatgccgttt
     3661 caatcataaa aacttttaag aacacatgac tggtaaaatt aatttaaaac aaatttaaat
     3721 tttcaacgta acattcaaca aaaatggtga aaactatcac ggaaattgtt aatattaata
     3781 tgtcccaaaa atagcctttg tatgtatatg atactaatcc atacatctat ggtatctata
     3841 ggtgagggcg atcgtaccaa agatggcaaa aagattggtt tgtggaaagt cttggccaat
     3901 cctgctatga tcatgttgat gattgctgct tccattcgtc actctggcgg catgactttc
     3961 gcttataatg ccgatttata ctacaacact tatttcccag atgttgatct tggctggtgg
     4021 ttgttcgctg tgaccattgg tattggtagt gttggtgtag ttgttggtgg tgttgtttct
     4081 gataaaattg tcgctaagat gggcataaga tcacgcgcat tggtacttgc tgtgagtcaa
     4141 ctgattgcca ctttgcctgc ttttggctcc gtatactatg atccactatg ggctatgatt
     4201 accttgggtt gtagttattt cttcgctgag atgtggttcg gcattgtgtt tgcgatcgtt
     4261 gtggaaattg taccgttgca agtgcgctct tcaacaattg gcgtattcct atttgtgatg
     4321 aacaacgttg gaggaaattt gccaattctc ttggatcctg tagcaaaaat gattggcttc
     4381 cgtgaggcgt ttatgatctt ttatgctggt ttctatggca taagtaagtc tgagttgttg
     4441 aaaaattcta tatatgtatg tatatacata tgtgcatgct ttatttattg tgtatattct
     4501 taaattctta ggctccgtgc tcttccttat cacctgcttc ggtacctcac gtaataagtg
     4561 tgcgttgaat ttattcacaa aaacattgca tattttcggc aaagtaaaat tttgttgcat
     4621 accttatcaa aaaataagtg ctgcatactt tttagagaaa ccaaataatt ttttattgca
     4681 tacccgtttt taataaaata cattgcatac cctcttttaa taaaaaatat tgcatacttt
     4741 gacgaaacaa attttcgttg catacccaat aaaagattat tatattgcat acccgttttt
     4801 aataaaatac attgcatacc ctcttttaat aaaaaatatt gcatacgttg acgaaacaaa
     4861 ttttcgttgc atacccaata aaagattatt atattgcata ccttttcttg ccataccatt
     4921 tagccgatca attccgcggc atactcggtg gcctccccac caccaacttt tttgcactgc
     4981 aaaaaaacac gcttttgcac gcgggcccat acatagtaca aactctacgt ttcgtagact
     5041 attttacata aatagtctac accgttgtat acgctccaaa tacactacca cacattgaac
     5101 ctttttgcag tgcaaaaaag tacgtgtcgg cagtcacgta ggccggcctt atcgggtcgc
     5161 gtcctgtcac gtacgaatca cattatcgga ccggacgagt gttgtcttat cgtgacagga
     5221 cgccagcttc ctgtgttgct aaccgcagcc ggacgcaact ccttatcgga acaggacgcg
     5281 cctccatatc agccgcgcgt tatctcatgc gcgtgaccgg acacgaggcg cccgtcccgc
     5341 ttatcgcgcc tataaataca gcccgcaacg atctggtaaa cacagttgaa cagcatctgt
     5401 tacagcgaca caacatgagc cggtccaaca acgccaacgc gcccacgcca tccaaccgcc
     5461 gccgcaacct gtctctggtg atggtgcgct cctccaagaa cgtcatcaag gagttcatgc
     5521 gcttcaaggt gcgcatggag ggcaccgtga acggccacga gttcgagatc gagggcgagg
     5581 gcgagggccg cccctacgag ggccacaaca ccgtgaagct gaaggtgacc aagggcggcc
     5641 ccctgccctt cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg
     5701 tgaagcaccc cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt
     5761 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     5821 tgcaggacgg ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc ccctccgacg
     5881 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     5941 gcgacggcgt gctgaagggc gagatccaca aggccctgaa gctgaaggac ggcggccact
     6001 acctggtgga gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg cccggctact
     6061 actacgtgga ctccaagctg gacatcacct cccacaacga ggactacacc atcgtggagc
     6121 agtacgagcg caccgagggc cgccaccacc tgttcctgta ggactctaga tcataatcag
     6181 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     6241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     6301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6361 tagttgtggt ttgtccaaac tcatcaatgt atcttaaagc ttgtcgacga tgtaggtcac
     6421 ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc
     6481 cacctcaccc atctggtcca tcatgatgaa cgggtcgagg tggcggtagt tgatcccggc
     6541 gaacgcgcgg cgcaccggga agccctcgcc ctcgaaaccg ctgggcgcgg tggtcacggt
     6601 gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg cggggcagcg tcagcgggtt
     6661 ctcgacggtc acggcgggca tgtcgacaga tatctataac aagaaaatat atatataata
     6721 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     6781 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     6841 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     6901 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     6961 tgcatgcgtc aattttacgc agactatctt tctagggtta atccctttag tgagggttaa
     7021 tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7081 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7141 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7201 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7261 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7321 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7381 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7441 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7501 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7561 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7621 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7681 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7741 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7801 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7861 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7921 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7981 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8041 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8101 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8161 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8221 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8281 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8341 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8401 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8461 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8521 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8581 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8641 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8701 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8761 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8821 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8881 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8941 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9001 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9061 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9121 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9181 gaaaagtgcc ac
//
