LOCUS       sequence-213959-        9040 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 9040)
  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 9040)
  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..9040
                     /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   3013..3075
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4796..5226
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     regulatory      5495..5504
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5634..5652)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5902..5921
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6096..6119
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    7053..7174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7090..7109)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7144..7163
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    7252..7351
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(8733..8749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8733..8749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8746..8768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8757..8773
                     /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(8781..8787,8788..8805,8806..8811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8826..8847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8964..8981)
                     /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 gctttctcat agctcacgct
      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 agaagaacag
      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 tttaaaaata taacaaaatt
     2341 gtgatcccac aaaatgaagt ggggcaaaat caaataatta atagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcc gtgactataa cacaagttga
     2461 tttaataatt ttagccaaca cgtcgggctg cgtgtttttt gccgacgcgt ctgtgtacac
     2521 gttgattaac tggtcgatta aactgttgaa ataatttaat ttttggttct tctttaaatc
     2581 tgtgatgaaa ttttttaaaa taactttaaa ttcttcattg gtaaaaaatg ccacgttttg
     2641 caacttgtga gggtctaata tgaggtcaaa ctcagtagga gttttatcca aaaaagaaaa
     2701 catgattacg tctgtacacg aacgcgtatt aacgcagagt gcaaagtata agagggttaa
     2761 aaaatatatt ttacgcacca tatacgcatc gggttgatat cgttaatatg gatcaatttg
     2821 aacagttgat taacgtgtct ctgctcaagt ctttgatcaa aacgcaaatc gacgaaaatg
     2881 tgtcggacaa tatcaagtcg atgagcgaaa aactaaaaag gctagaatac gacaatctca
     2941 cagacagcgt tgagatatac ggtattcacg acagcaggct gaataataaa aaaattagaa
     3001 actattattt aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgcg
     3061 taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata
     3121 gatattaagt tttattatat ttacacttac atactaataa taaattcaac aaacaattta
     3181 tttatgttta tttatttatt aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac
     3241 aaaactttta aacattctct cttttacaaa aataaactta ttttgtactt taaaaacagt
     3301 catgttgtat tataaaataa gtaattagct taacttatac ataatagaaa caaattatac
     3361 ttattagtca gtcagaaaca actttggcac atatcaatat tatgctctcg acaaataact
     3421 tttttgcatt ttttgcacga tgcatttgcc tttcgcctta ttttagaggg gcagtaagta
     3481 cagtaagtac gttttttcat tactggctct tcagtactgt catctgatgt accaggcact
     3541 tcatttggca aaatattaga gatattatcg cgcaaatatc tcttcaaagt aggagcttct
     3601 aaacgcttac gcataaacga tgacgtcagg ctcatgtaaa ggtttctcat aaattttttg
     3661 cgactttgaa ccttttctcc cttgctactg acattatggc tgtatataat aaaagaattt
     3721 atgcaggcaa tgtttatcat tccgtacaat aatgccatag gccacctatt cgtcttccta
     3781 ctgcagggga agcttcatgg gatctgctgg ccattatgta ggcgctcccg gggaaacata
     3841 gttaagtttg tgttgcggtt tgatgattgc gttgtacagc aacaaacctt tcaccaaacg
     3901 agtgaaaagt gttgtgaatt tttttctcca acgccaacgc ttaactatac tctataacta
     3961 ctcattcacc aaaatctatt tgctctatcc ataaactagt tctttccaag taacttaatc
     4021 tcccgaatcg taatttctca ctcgaactat ggcgcgcata gttgtgctta aggttggagg
     4081 tgaggtgttt tgtagatgcc cgaagtccac ctgtggcttg ttcggtgaca aaaactgatc
     4141 tgattctata ttctagctct ggtctcaatt agacctccgc tcagagaagt aaataggaaa
     4201 tagtgtctta aatgctacat ttgtaagtgt taattcataa gtgctgcgtc gcacttattc
     4261 cgaattgctg tgttgcctac tcttgagtgg atcgattctt tgctgagtcg ttctcctatt
     4321 acatagtccg gaggagtagg ggtttcctgt ccaaatgtca tgttaacggc cgttcgctga
     4381 acggtcgttg tgtggcctaa ttaaatccgt tgtttacatt ttggttattt ggcactactt
     4441 ctagggtaag ccggcgtggc ttagctagag ggtatgtgtg ccgtttggaa ccggtaactc
     4501 ctccctcctt ggattgaagt gacagccaca acgaactagt agtcttcact aagctcggct
     4561 ctggtaccgt tttaaatgcc actcttcttc ttctattgat gaggtatatt gtcgtcacta
     4621 ctccaactaa tagtaatacg taaagtgctc cgaaatttgt agtcgtaaat tttttccaat
     4681 atatgatgga attagaagca tggttggatc ttatgttcag cttatgcagg ttaatgtctt
     4741 ctgtgttctc ggtgaaatat atatttttta taattgggga ttgtgcgttt aaacttggcc
     4801 acgtaataag tgtgcgttga atttattcgc aaaaacattg catattttcg gcaaagtaaa
     4861 attttgttgc ataccttatc aaaaaataag tgctgcatac tttttagaga aaccaaataa
     4921 ttttttattg catacccgtt tttaataaaa tacattgcat accctctttt aataaaaaat
     4981 attgcatact ttgacgaaac aaattttcgt tgcataccca ataaaagatt attatattgc
     5041 atacccgttt ttaataaaat acattgcata ccctctttta ataaagaata ttgcatacgt
     5101 tgacgaaaca aattttcgtt gcatacccaa taaaagatta ttatattgca taccttttct
     5161 tgccatacca tttagccgat caattgtgct cggcaacagt atatttgtgg tgtgccaacc
     5221 aacaacggcg cgcctaaatt cgagctcgcc cggggatcta attcaattag agactaattc
     5281 aattagagct aattcaatta ggatccaagc ttatcgattt cgaaccctcg accgccggag
     5341 tataaataga ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca
     5401 cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatcggg
     5461 gtaccgctag agtcgacggt acgatccacc ggtcgccacc atggtgagca agggcgagga
     5521 ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     5581 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     5641 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     5701 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     5761 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     5821 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     5881 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     5941 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     6001 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     6061 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     6121 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctggg
     6181 gcatggcacc ggcagcaccg gcagcggcag ctccggcacc gcctcctccg aggacaacaa
     6241 catggccgtt atcaaggaat ttatgcgctt caaagttagg atggagggat ccatgaacgg
     6301 acatgagttc gagatcgagg gagagggcga gggacgcccg tatgaaggca cacaaacagc
     6361 caaactcaag gtcaccaagg gcggaccact gcccttcgcc tgggatatcc tgagtcccca
     6421 gtttatgtac ggcagcaagg cctacgttaa gcaccccgct gacataccgg actacaaaaa
     6481 gctgtccttt ccggaaggct tcaagtggga gcgcgtgatg aatttcgaag acggaggact
     6541 ggtcactgtg acccaagata gcagtttgca ggacggtaca ctgatctata aggttaaaat
     6601 gcgcggcact aactttccgc cagatggccc agtgatgcag aagaagacca tgggttggga
     6661 ggcatccacc gaacgtctgt accctcgaga cggagtgctc aagggcgaga tccatcaggc
     6721 cctcaaactg aaagatggtg gtcactacct ggtcgaattt aagaccattt acatggccaa
     6781 gaagccggtt cagctgcccg gatattatta tgtggatacg aaactggata taacttcgca
     6841 taacgaagac tacaccattg tcgagcagta tgagcgcagc gaaggccgac atcacctgtt
     6901 cctctacggc atggacgagc tgtacaagta ggcggccgcg actctagatc ataatcagcc
     6961 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     7021 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     7081 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7141 gttgtggttt gtccaaactc atcaatgtat cttaaagctt cctagggtac tgacggacac
     7201 accgaagccc cggcggcaac cctcagcgga tgccccgggg cttcacgttt tcccaggtca
     7261 gaagcggttt tcgggagtag tgccccaact ggggtaacct ttgagttctc tcagttgggg
     7321 gcgtagggtc gccgacatga cacaaggggt tgtgaccggg gtggacacgt acgcgggtgc
     7381 ttacgaccgt cagtcgcgcg agcgcgaggg cccgggtttt ttaaatggta cccttgcagc
     7441 gccacctggc cgcgaagagt tccattctct gcttcgaagt acgaagagag aaagagtact
     7501 ttaaatttcc acattcccgc cttgcagcgc cacctggcct tggtaatgta gaactaggaa
     7561 ggaaagcacc aacacaagat gtcgctctcc gacgatggac atgtcgcgta aaaaatgttc
     7621 gataactttc aatggttcga ttgaacagac aataagtgta tttaagacac cagttcttat
     7681 attcaaaaat cctaacaact cacaccttgc agcgccacct ggccctagag atccaccaac
     7741 tctagagaga tcttacaatt tgattcaacg gtagtgtttt gtgtgtttaa tattaaaatg
     7801 tgagctggtt ctacttcttt tataatttct tgattcccta tgtctttagt tttaaaattg
     7861 gcagtttcca tcttatgtat gttattgaga cattcagttt taatattacc tctgattgcc
     7921 attgcattgt cgcagatatg gtatttatat atcttgttat atttgttgga aaagtagtaa
     7981 aaaacatgcc atatagttag gtatatttat atatttatat ttatatttca tttatgggta
     8041 atgggacaat gtgtgtgaac ttttagtaat tttgtttgaa tgcagggatt cttattctaa
     8101 agtttatctt aattccaatc atgaaagtat ctaattctaa catttgaaat atgtgtttgt
     8161 ctgacatttc ccgtaactcc gggtcgttta attcttgtct gattacgttt acttcctcct
     8221 cagatacaat aaaagtgggt acaatcttta gtttggctag taacattttt aaaatagttc
     8281 gacaaaacat ccaaatacat gttaattcta cttgcttact taattgactt gctcgaaaat
     8341 gggcagtata attatttaag gcaagttcgc gtccaactgg aatgaatcct gtttgttttt
     8401 cgaggcttct gaccgaaact ttcctccttc aacgaaatta tattctgctt cagtatcatt
     8461 tgtcttgttg tttgtctcga aattaaacaa tctcagtccg tttgtaattc aatctactag
     8521 gattatcatc gcttggcaaa atggcagaat atctcgaatt tcggttgttg ttatcaactg
     8581 atactatatt gatactatac tgatattgat tgtaacttgg gcgaaagcaa tcgttctgcc
     8641 tcatttgttg tctgctttga ctgactgaaa aaagcgtaaa gtgcagcccc cgcagaacac
     8701 acgatgggac tgtttcagaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     8761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     8821 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     8881 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     8941 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     9001 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
//
