LOCUS       sequence-208913-        9026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates marker gene onto D. melanogaster Y chromosome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9026)
  AUTHORS   Buchman A, Akbari OS
  TITLE     Site-specific transgenesis of the D. melanogaster Y-chromosome using
            CRISPR/Cas9.
  JOURNAL   Insect Mol Biol. 2018 Aug 6. doi: 10.1111/imb.12528.
  PUBMED    30079589
REFERENCE   2  (bases 1 to 9026)
  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: "Insect Mol 
            Biol. 2018 Aug 6. doi: 10.1111/imb.12528."
COMMENT     Sequence Label: AByF
FEATURES             Location/Qualifiers
     source          1..9026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(95..683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(175..194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(854..1645,1646..1714))
                     /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     1477..1496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1715..1819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1887..1905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1943..1965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2065..2084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2278..2300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2292..2309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2293..2309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2998..3060
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4781..5211
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     regulatory      5480..5489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5619..5637)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5887..5906
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6081..6104
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    7038..7159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7075..7094)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7129..7148
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    7237..7336
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(8719..8735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8719..8735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8732..8754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8743..8759
                     /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(8767..8773,8774..8791,8792..8797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8812..8833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8950..8967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
       61 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
      301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2221 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2281 agtcacgacg ttgtaaaacg acggccagtg ccaagctttg tttaaaatat aacaaaattg
     2341 tgatcccaca aaatgaagtg gggcaaaatc aaataattaa ctagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcg tgactataac acaagttgat
     2461 ttaataattt tagccaacac gtcgggctgc gtgttttttg cgctctgtgt acacgttgat
     2521 taactggtcg attaaataat ttaatttttg gttcttcttt aaatctgtga tgaaattttt
     2581 taaaataact ttaaattctt cattggtaaa aaatgccacg ttttgcaact tgtgagggtc
     2641 taatatgagg tcaaactcag taggagtttt atccaaaaaa gaaaacatga ttacgtctgt
     2701 acacgaacgc gtattaacgc agagtgcaaa gtataagagg gttaaaaaat atattttacg
     2761 caccatatac gcatcgggtt gatatcgtta atatggatca atttgaacag ttgattaacg
     2821 tgtctctgct caagtctttg atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca
     2881 agtcgatgag cgaaaaacta aaaaggctag aatacgacaa tctcacagac agcgttgaga
     2941 tatacggtat tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc
     3001 tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg
     3061 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
     3121 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
     3181 ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat
     3241 tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa
     3301 aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt agtcagtcag
     3361 aaacaacttt ggcacatatc aatattatgc tctcgacaaa taactttttt gcattttttg
     3421 cacgatgcat ttgcctttcg ccttatttta gaggggcagt aagtacagta agtacgtttt
     3481 ttcattactg gctcttcagt actgtcatct gatgtaccag gcacttcatt tggcaaaata
     3541 ttagagatat tatcgcgcaa atatctcttc aaagtaggag cttctaaacg cttacgcata
     3601 aacgatgacg tcaggctcat gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt
     3661 tctcccttgc tactgacatt atggctgtat ataataaaag aatttatgca ggcaatgttt
     3721 atcattccgt acaataatgc cataggccac ctattcgtct tcctactgca ggggaagctt
     3781 catgggatct gctggccatt atgtaggcgc tcccggggaa acatagttaa gtttgtgttg
     3841 cggtttgatg attgcgttgt acagcaacaa acctttcacc aaacgagtga aaagtgttgt
     3901 gaattttttt ctccaacgcc aacgcttaac tatactctat aactactcat tcaccaaaat
     3961 ctatttgctc tatccataaa ctagttcttt ccaagtaact taatctcccg aatcgtaatt
     4021 tctcactcga actatggcgc gcatagttgt gcttaaggtt ggaggtgagg tgttttgtag
     4081 atgcccgaag tccacctgtg gcttgttcgg tgacaaaaac tgatctgatt ctatattcta
     4141 gctctggtct caattagacc tccgctcaga gaagtaaata ggaaatagtg tcttaaatgc
     4201 tacatttgta agtgttaatt cataagtgct gcgtcgcact tattccgaat tgctgtgttg
     4261 cctactcttg agtggatcga ttctttgctg agtcgttctc ctattacata gtccggagga
     4321 gtaggggttt cctgtccaaa tgtcatgtta acggccgttc gctgaacggt cgttgtgtgg
     4381 cctaattaaa tccgttgttt acattttggt tatttggcac tacttctagg gtaagccggc
     4441 gtggcttagc tagagggtat gtgtgccgtt tggaaccggt aactcctccc tccttggatt
     4501 gaagtgacag ccacaacgaa ctagtagtct tcactaagct cggctctggt accgttttaa
     4561 atgccactct tcttcttcta ttgatgaggt atattgtcgt cactactcca actaatagta
     4621 atacgtaaag tgctccgaaa tttgtagtcg taaatttttt ccaatatatg atggaattag
     4681 aagcatggtt ggatcttatg ttcagcttat gcaggttaat gtcttctgtg ttctcggtga
     4741 aatatatatt ttttataatt ggggattgtg cgtttaaact tggccacgta ataagtgtgc
     4801 gttgaattta ttcgcaaaaa cattgcatat tttcggcaaa gtaaaatttt gttgcatacc
     4861 ttatcaaaaa ataagtgctg catacttttt agagaaacca aataattttt tattgcatac
     4921 ccgtttttaa taaaatacat tgcataccct cttttaataa aaaatattgc atactttgac
     4981 gaaacaaatt ttcgttgcat acccaataaa agattattat attgcatacc cgtttttaat
     5041 aaaatacatt gcataccctc ttttaataaa aaatattgca tacgttgacg aaacaaattt
     5101 tcgttgcata cccaataaaa gattattata ttgcatacct tttcttgcca taccatttag
     5161 ccgatcaatt gtgctcggca acagtatatt tgtggtgtgc caaccaacaa cggcgcgcct
     5221 aaattcgagc tcgcccgggg atctaattca attagagact aattcaatta gagctaattc
     5281 aattaggatc caagcttatc gatttcgaac cctcgaccgc cggagtataa atagaggcgc
     5341 ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa
     5401 gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tcggggtacc gctagagtcg
     5461 acggtacgat ccaccggtcg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt
     5521 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
     5581 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5641 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
     5701 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
     5761 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     5821 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     5881 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     5941 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     6001 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     6061 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     6121 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag
     6181 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgttatcaa
     6241 ggaatttatg cgcttcaaag ttaggatgga gggatccatg aacggacatg agttcgagat
     6301 cgagggagag ggcgagggac gcccgtatga aggcacacaa acagccaaac tcaaggtcac
     6361 caagggcgga ccactgccct tcgcctggga tatcctgagt ccccagttta tgtacggcag
     6421 caaggcctac gttaagcacc ccgctgacat accggactac aaaaagctgt cctttccgga
     6481 aggcttcaag tgggagcgcg tgatgaattt cgaagacgga ggactggtca ctgtgaccca
     6541 agatagcagt ttgcaggacg gtacactgat ctataaggtt aaaatgcgcg gcactaactt
     6601 tccgccagat ggcccagtga tgcagaagaa gaccatgggt tgggaggcat ccaccgaacg
     6661 tctgtaccct cgagacggag tgctcaaggg cgagatccat caggccctca aactgaaaga
     6721 tggtggtcac tacctggtcg aatttaagac catttacatg gccaagaagc cggttcagct
     6781 gcccggatat tattatgtgg atacgaaact ggatataact tcgcataacg aagactacac
     6841 cattgtcgag cagtatgagc gcagcgaagg ccgacatcac ctgttcctct acggcatgga
     6901 cgagctgtac aagtaggcgg ccgcgactct agatcataat cagccatacc acatttgtag
     6961 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     7021 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     7081 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     7141 aactcatcaa tgtatcttaa agcttcctag ggtactgacg gacacaccga agccccggcg
     7201 gcaaccctca gcggatgccc cggggcttca cgttttccca ggtcagaagc ggttttcggg
     7261 agtagtgccc caactggggt aacctttgag ttctctcagt tgggggcgta gggccgccga
     7321 catgacacaa ggggttgtga ccggggtgga cacgtacgcg ggtgcttacg accgtcagtc
     7381 gcgcgagcgc gagggcccgg gttttttaaa tggtacccct tgcagcgcca cctggccgcg
     7441 aagagttcca ttctctgctt cgaagtacga agagagaaag agtactttaa atttccacat
     7501 tcccgccttg cagcgccacc tggccttggt aatgtagaac taggaaggaa agcaccaaca
     7561 caagatgtcg ctctccgacg atggacatgt cgcgtaaaaa atgttcgata actttcaatg
     7621 gttcgattga acagacaata agtgtattta agacaccagt tcttatattc aaaaatccta
     7681 acaactcaca ccttgcagcg ccacctggcc ctagagatcc accaactcta gagagatctt
     7741 acaatttgat tcaacggtag tgttttgtgt gtttaatatt aaaatgtgag ctggttctac
     7801 ttcttttata atttcttgat tccctatgtc tttagtttta aaattggcag tttccatctt
     7861 atgtatgtta ttgagacatt cagttttaat attacctctg attgccattg cattgtcgca
     7921 gatatggtat ttatatatct tgttatattt gttggaaaag tagtaaaaaa catgccatat
     7981 agttaggtat atttatatat ttatatttat atttcattta tgggtaatgg gacaatgtgt
     8041 gtgaactttt agtaattttg tttgaatgca gggattctta ttctaaagtt tatcttaatt
     8101 ccaatcatga aagtatctaa ttctaacatt tgaaatatgt gtttgtctga catttcccgt
     8161 aactccgggt cgtttaattc ttgtctgatt acgtttactt cctcctcaga tacaataaaa
     8221 gtgggtacaa tctttagttt ggctagtaac atttttaaaa tagttcgaca aaacatccaa
     8281 atacatgtta attctacttg cttacttaat tgacttgctc gaaaatgggc agtataatta
     8341 tttaaggcaa gttcgcgtcc aactggaatg aatcctgttt gtttttcgag gcttctgacc
     8401 gaaactttcc tccttcaacg aaattatatt ctgcttcagt atcatttgtc ttgttgtttg
     8461 tctcgaaatt aaacaatctc agtccgtttg taattcaatc tactaggatt atcatcgctt
     8521 ggcaaaatgg cagaatatct cgaatttcgg ttgttgttat caactgatac tatattgata
     8581 ctatactgat attgattgta acttgggcga aagcaatcgt tctgcctcat ttgttgtctg
     8641 ctttgactga ctgaaaaaag cgtaaagtgc agcccccgca gaacacacga tgggactgtt
     8701 tcagaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     8761 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     8821 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8881 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8941 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     9001 agctcactca aaggcggtaa tacggt
//
