LOCUS       sequence-213958-        8700 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 8700)
  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 8700)
  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: AByG
FEATURES             Location/Qualifiers
     source          1..8700
                     /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    4608..5038
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     regulatory      5307..5316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5446..5464)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5714..5733
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5908..5931
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    6865..6986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6902..6921)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6956..6975
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    7064..7163
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(8393..8409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8393..8409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8406..8428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8417..8433
                     /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(8441..8447,8448..8465,8466..8471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8486..8507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8624..8641)
                     /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 ctgcaggtgt acctcgccca gttggtccta ttggttgagg gcctacacgg tcgctcgatg
     3841 tgttgtagtt ggtgcaaaag atgaaagatt accggacagt gaccagatga gagttctgga
     3901 agtgcctcag ctgtaatcat tgattgggat atcctttttg tgatggcaaa gtcaatcaaa
     3961 tctggaagct tcatgggatc tgctggccat tatgtaggcg ctcccgggga aacatagtta
     4021 agtttgtgtt gcggtttgat gattgcgttg tacagcaaca aacctttcac caaacgagtg
     4081 aaaagtgttg tgaatttttt tctccaacgc caacgcttaa ctatactcta taactactca
     4141 ttcaccaaaa tctatttgct ctatccataa actagttctt tccaagtaac ttaatctccc
     4201 gaatcgtaat ttctcactcg aactatggcg cgcatagttg tgcttaaggt tggaggtgag
     4261 gtgttttgta gatgcccgaa gtccacctgt ggcttgttcg gtgacaaaaa ctgatctgat
     4321 tctatattct agctctggtc tcaattagac ctccgctcag agaagtaaat aggaaatagt
     4381 gtcttaaatg ctacatttgt aagtgttaat tcataagtgc tgcgtcgcac ttattccgaa
     4441 ttgctgtgtt gcctactctt gagtggatcg attctttgct gagtcgttct cctattacat
     4501 agtccggagg agtaggggtt tcctgtccaa atgtcatgtt aacggccgtt cgctgaacgg
     4561 tcgttgtgtg gcctaattaa atccgttgtt tacattttgt ttaaacttgg ccacgtaata
     4621 agtgtgcgtt gaatttattc gcaaaaacat tgcatatttt cggcaaagta aaattttgtt
     4681 gcatacctta tcaaaaaata agtgctgcat actttttaga gaaaccaaat aattttttat
     4741 tgcatacccg tttttaataa aatacattgc ataccctctt ttaataaaaa atattgcata
     4801 ctttgacgaa acaaattttc gttgcatacc caataaaaga ttattatatt gcatacccgt
     4861 ttttaataaa atacattgca taccctcttt taataaagaa tattgcatac gttgacgaaa
     4921 caaattttcg ttgcataccc aataaaagat tattatattg catacctttt cttgccatac
     4981 catttagccg atcaattgtg ctcggcaaca gtatatttgt ggtgtgccaa ccaacaacgg
     5041 cgcgcctaaa ttcgagctcg cccggggatc taattcaatt agagactaat tcaattagag
     5101 ctaattcaat taggatccaa gcttatcgat ttcgaaccct cgaccgccgg agtataaata
     5161 gaggcgcttc gtctacggag cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta
     5221 agcgaaagct aagcaaataa acaagcgcag ctgaacaagc taaacaatcg gggtaccgct
     5281 agagtcgacg gtacgatcca ccggtcgcca ccatggtgag caagggcgag gaggtcatca
     5341 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
     5401 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     5461 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     5521 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     5581 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     5641 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     5701 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     5761 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     5821 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     5881 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     5941 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg gggcatggca
     6001 ccggcagcac cggcagcggc agctccggca ccgcctcctc cgaggacaac aacatggccg
     6061 ttatcaagga atttatgcgc ttcaaagtta ggatggaggg atccatgaac ggacatgagt
     6121 tcgagatcga gggagagggc gagggacgcc cgtatgaagg cacacaaaca gccaaactca
     6181 aggtcaccaa gggcggacca ctgcccttcg cctgggatat cctgagtccc cagtttatgt
     6241 acggcagcaa ggcctacgtt aagcaccccg ctgacatacc ggactacaaa aagctgtcct
     6301 ttccggaagg cttcaagtgg gagcgcgtga tgaatttcga agacggagga ctggtcactg
     6361 tgacccaaga tagcagtttg caggacggta cactgatcta taaggttaaa atgcgcggca
     6421 ctaactttcc gccagatggc ccagtgatgc agaagaagac catgggttgg gaggcatcca
     6481 ccgaacgtct gtaccctcga gacggagtgc tcaagggcga gatccatcag gccctcaaac
     6541 tgaaagatgg tggtcactac ctggtcgaat ttaagaccat ttacatggcc aagaagccgg
     6601 ttcagctgcc cggatattat tatgtggata cgaaactgga tataacttcg cataacgaag
     6661 actacaccat tgtcgagcag tatgagcgca gcgaaggccg acatcacctg ttcctctacg
     6721 gcatggacga gctgtacaag taggcggccg cgactctaga tcataatcag ccataccaca
     6781 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     6841 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     6901 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6961 ttgtccaaac tcatcaatgt atcttaaagc ttcctagggt actgacggac acaccgaagc
     7021 cccggcggca accctcagcg gatgccccgg ggcttcacgt tttcccaggt cagaagcggt
     7081 tttcgggagt agtgccccaa ctggggtaac ctttgagttc tctcagttgg gggcgtaggg
     7141 tcgccgacat gacacaaggg gttgtgaccg gggtggacac gtacgcgggt gcttacgacc
     7201 gtcagtcgcg cgagcgcgag ggcccgggtt ttttaaatgg tacccttgca gcgccacctg
     7261 gccgcgaaga gttccattct ctgcttcgaa gtacgaagag agaaagagta ctttaaattt
     7321 ccacattccc gccttgcagc gccacctggc cttggtaatg tagaactagg aaggaaagca
     7381 ccaacacaag atgtcgctct ccgacgatgg acatgtcgcg taaaaaatgt tcgataactt
     7441 tcaatggttc gattgaacag acaataagtg tatttaagac accagttctt atattcaaaa
     7501 atcctaacaa ctcacacctt gcagcgccac ctggccctag agaccaccaa ctctagagag
     7561 atctgtaagc cggcgtggct tagctagagg gtatgtgtgc cgtttggaac cggtaactcc
     7621 tccctccttg gattgaagtg acagccacaa cgaactagta gtcttcacta agctcggctc
     7681 tggtaccgtt ttaaatgcca ctcttcttct tctattgatg aggtatattg tcgtcactac
     7741 tccaactaat agtaatacgt aaagtgctcc gaaatttgta gtcgtaaatt ttttccaata
     7801 tatgatggaa ttagaagcat ggttggatct tatgttcagc ttatgcaggt taatgtcttc
     7861 tgtgttctcg gtgaaatata tattttttat aattggggat tgtgcggatc ttcttatgga
     7921 aattttggta caatttgatt caacggtagt gttttgtgtg tttaatatta aaatgtgagc
     7981 tggttctact tcttttataa tttcttgatt ccctatgtct ttagttttaa aattggcagt
     8041 ttccatctta tgtatgttat tgagacattc agttttaata ttacctctga ttgccattgc
     8101 attgtcgcag atatggtatt tatatatctt gttatatttg ttggaaaagt agtaaaaaac
     8161 atgccatata gttaggtata tttatatatt tatatttata tttcatttat gggtaatggg
     8221 acaatgtgtg tgaactttta gtaattttgt ttgaatgcag ggattcttat tctaaagttt
     8281 atcttaattc caatcatgaa agtatctaat tctaacattt gaaatatgtg tttgtctgac
     8341 atttcccgta actccgggtc gtttaattct tgtctgagaa ttcgtaatca tggtcatagc
     8401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     8461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     8521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     8581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     8641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
//
