LOCUS       sequence-327698-        9202 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 9202)
  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 9202)
  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: AByH
FEATURES             Location/Qualifiers
     source          1..9202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..431
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     regulatory      700..709
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(839..857)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1107..1126
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1301..1324
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2258..2379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2295..2314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2349..2368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2457..2556
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(3988..4004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3988..4004)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4001..4023)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4012..4028
                     /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(4036..4042,4043..4060,4061..4066))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4081..4102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4219..4236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4390..4978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4470..4489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5149..5940,5941..6009))
                     /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     5772..5791
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6010..6114)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6182..6200
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6238..6260)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6360..6379
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6573..6595
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6587..6604
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6588..6604
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7308..7370
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 tggccacgta ataagtgtgc gttgaattta ttcgcaaaaa cattgcatat tttcggcaaa
       61 gtaaaatttt gttgcatacc ttatcaaaaa ataagtgctg catacttttt agagaaacca
      121 aataattttt tattgcatac ccgtttttaa taaaatacat tgcataccct cttttaataa
      181 aaaatattgc atactttgac gaaacaaatt ttcgttgcat acccaataaa agattattat
      241 attgcatacc cgtttttaat aaaatacatt gcataccctc ttttaataaa gaatattgca
      301 tacgttgacg aaacaaattt tcgttgcata cccaataaaa gattattata ttgcatacct
      361 tttcttgcca taccatttag ccgatcaatt gtgctcggca acagtatatt tgtggtgtgc
      421 caaccaacaa cggcgcgcct aaattcgagc tcgcccgggg atctaattca attagagact
      481 aattcaatta gagctaattc aattaggatc caagcttatc gatttcgaac cctcgaccgc
      541 cggagtataa atagaggcgc ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt
      601 gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg cagctgaaca agctaaacaa
      661 tcggggtacc gctagagtcg acggtacgat ccaccggtcg ccaccatggt gagcaagggc
      721 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
      781 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
      841 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
      901 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
      961 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     1021 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     1081 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1141 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     1201 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     1261 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     1321 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     1381 ctggggcatg gcaccggcag caccggcagc ggcagctccg gcaccgcctc ctccgaggac
     1441 aacaacatgg ccgttatcaa ggaatttatg cgcttcaaag ttaggatgga gggatccatg
     1501 aacggacatg agttcgagat cgagggagag ggcgagggac gcccgtatga aggcacacaa
     1561 acagccaaac tcaaggtcac caagggcgga ccactgccct tcgcctggga tatcctgagt
     1621 ccccagttta tgtacggcag caaggcctac gttaagcacc ccgctgacat accggactac
     1681 aaaaagctgt cctttccgga aggcttcaag tgggagcgcg tgatgaattt cgaagacgga
     1741 ggactggtca ctgtgaccca agatagcagt ttgcaggacg gtacactgat ctataaggtt
     1801 aaaatgcgcg gcactaactt tccgccagat ggcccagtga tgcagaagaa gaccatgggt
     1861 tgggaggcat ccaccgaacg tctgtaccct cgagacggag tgctcaaggg cgagatccat
     1921 caggccctca aactgaaaga tggtggtcac tacctggtcg aatttaagac catttacatg
     1981 gccaagaagc cggttcagct gcccggatat tattatgtgg atacgaaact ggatataact
     2041 tcgcataacg aagactacac cattgtcgag cagtatgagc gcagcgaagg ccgacatcac
     2101 ctgttcctct acggcatgga cgagctgtac aagtaggcgg ccgcgactct agatcataat
     2161 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2221 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2281 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2341 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa agcttcctag ggtactgacg
     2401 gacacaccga agccccggcg gcaaccctca gcggatgccc cggggcttca cgttttccca
     2461 ggtcagaagc ggttttcggg agtagtgccc caactggggt aacctttgag ttctctcagt
     2521 tgggggcgta gggtcgccga catgacacaa ggggttgtga ccggggtgga cacgtacgcg
     2581 ggtgcttacg accgtcagtc gcgcgagcgc gagggcccgg gttttttaaa tggtaccctt
     2641 gcagcgccac ctggccgcga agagttccat tctctgcttc gaagtacgaa gagagaaaga
     2701 gtactttaaa tttccacatt cccgccttgc agcgccacct ggccttggta atgtagaact
     2761 aggaaggaaa gcaccaacac aagatgtcgc tctccgacga tggacatgtc gcgtaaaaaa
     2821 tgttcgataa ctttcaatgg ttcgattgaa cagacaataa gtgtatttaa gacaccagtt
     2881 cttatattca aaaatcctaa caactcacac cttgcagcgc cacctggccc tagagatcca
     2941 ccaactctag agagatcttg cgaacagcat taagtacatg catcaacgat taccaggata
     3001 tgctcattac gcttgtaact tctcggaaaa ggtcctagat gatagatgtg taccgtagcg
     3061 aacggtactg gcttgatgtt atcgtaatgg tattgaccct cttggcgacc tcctcgttgc
     3121 ttatacaggg aacaacctac gcatgaatta atgtatcctt taacataatt tcgcatacag
     3181 gggaaccaga attgatttat aattcggctg attgttttgt cggtgacatt ctacaagaat
     3241 gcgatagcgt aactgctttg gtacaaccca aaggtgcttg ttgttatgtt tgcgatataa
     3301 gcgaccaagt tcaaggacat actcatcgtt ctcagtgatc ttcttagcaa tcatttcgct
     3361 agcaattcgt tttattttct cgtcctgcaa ttgtacagca aacaaccaat cggcttcatc
     3421 aatgacggtt ttgaatgtct ttaggtcagc ttcttccatt tcacgcgaat cttcatatgg
     3481 tgatcggctt aaggcatcga cgtgttccat acacgtaact gagcgatgca gtatctcaat
     3541 atcgaattct tggatgcgta gccaccatct ggcaattcgg ggtatcagct ctcgtttctc
     3601 catcgtcgtt cttaacgcat tacaatcagt gatgactcga attcgtttcc catagacata
     3661 atacttgaaa cgttccagtg actcgacaac agccaaagct tctagctcgt agatgtgaaa
     3721 cttcatttcg ctgtatgtcg tcgatcggct ataataggct actggcttta actgatcatg
     3781 ttcctttgca atagaactcc ggctaaacaa acagaactag catcagtatg tatttcgtgt
     3841 tctgggctga cgtcgtagct aataagaaca ggcttcgaaa taaggagttg tttaagttca
     3901 ttgaatgcgt catcttgctg tggttcccat ttaaacggct ggtctttgcg tagtagcttg
     3961 cgtaaggcct cggaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4021 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4081 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4141 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4201 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4801 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6001 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6121 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6181 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6241 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6301 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6361 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6421 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6481 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6541 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     6601 cagtgccaag ctttgtttaa aaatataaca aaattgtgat cccacaaaat gaagtggggc
     6661 aaaatcaaat aattaatagt gtccgtaaac ttgttggtct tcaacttttt gaggaacacg
     6721 ttggacggca aatccgtgac tataacacaa gttgatttaa taattttagc caacacgtcg
     6781 ggctgcgtgt tttttgccga cgcgtctgtg tacacgttga ttaactggtc gattaaactg
     6841 ttgaaataat ttaatttttg gttcttcttt aaatctgtga tgaaattttt taaaataact
     6901 ttaaattctt cattggtaaa aaatgccacg ttttgcaact tgtgagggtc taatatgagg
     6961 tcaaactcag taggagtttt atccaaaaaa gaaaacatga ttacgtctgt acacgaacgc
     7021 gtattaacgc agagtgcaaa gtataagagg gttaaaaaat atattttacg caccatatac
     7081 gcatcgggtt gatatcgtta atatggatca atttgaacag ttgattaacg tgtctctgct
     7141 caagtctttg atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca agtcgatgag
     7201 cgaaaaacta aaaaggctag aatacgacaa tctcacagac agcgttgaga tatacggtat
     7261 tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc tagaaagata
     7321 atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg
     7381 gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca
     7441 cttacatact aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa
     7501 aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat tctctctttt
     7561 acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa aataagtaat
     7621 tagcttaact tatacataat agaaacaaat tatacttatt agtcagtcag aaacaacttt
     7681 ggcacatatc aatattatgc tctcgacaaa taactttttt gcattttttg cacgatgcat
     7741 ttgcctttcg ccttatttta gaggggcagt aagtacagta agtacgtttt ttcattactg
     7801 gctcttcagt actgtcatct gatgtaccag gcacttcatt tggcaaaata ttagagatat
     7861 tatcgcgcaa atatctcttc aaagtaggag cttctaaacg cttacgcata aacgatgacg
     7921 tcaggctcat gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt tctcccttgc
     7981 tactgacatt atggctgtat ataataaaag aatttatgca ggcaatgttt atcattccgt
     8041 acaataatgc cataggccac ctattcgtct tcctactgca gggacggaat taacatcgtc
     8101 ggcttgttca ggatttttcg atcccaaatt gatatatttt ttattagcgg ataaatctac
     8161 gctggcgtgt tcaggatttt cagatcccga actattagtt ttattaatga cggagaattc
     8221 aacgtcggcg tgctcaggat tgtcagatcc cgaattatat aaagtgcctt cagcaataac
     8281 catttcggtt ttaaatacct tttgctcatc aaagtatttt ttgataatat attgtaaggg
     8341 ggaatggctt tagtggtggt gaggtgagtg gtgaggcaat gagatatgag gcaaagcaaa
     8401 tataagttta ttaaatgaaa ttagatacag gaaatattaa ctgaattaaa tgtgatcact
     8461 gcgcgacggc ttgctcttcg acgacgaccg agcgaacaca cattaacggg gccatctgtt
     8521 ctcccgggtt accaccgtac ttgacaggca aggttgccat atgcttacag tcctgacgta
     8581 tcgtagcgac ctcgcacgaa tcattaagat atttatctgc tgggttgtcc ttatcatcgg
     8641 tgtcatctaa tggtatctgg caccacggct tgagtttatc ggacacatac acggatttat
     8701 agtgtcgctg agttcgttgg gcattcggca aatcttcgac caagtatcga tcatgtggta
     8761 gtttctttgc gatgattaac gggcctttat aacgaggctc gattttacgt gaagtgcctg
     8821 tactaccttg ttcgttttcg gccaaaacta agtctccttc gctgtaaacc gtcggcttgg
     8881 cgtgtttggc atcgtaacgt ttctttgctt ctgctcgatg ttcgtcaatt cgtgttgcag
     8941 tatcggtacg aagctgttgc aactcttcgg tagtcagcac ttggtcatca gtacattgca
     9001 acgctcgtac tacggagttc ttcaacgagt gtcgtggctc ttaacaatat aatagctgaa
     9061 atggagaacg attagtagtc ttgttggtca tcgtgttaat gctccactgg atttggcata
     9121 gtttctcatc ccagcgcttt tcattctcaa cagtaggcaa tagcattgat aggattatgc
     9181 ggttagtgcg tttgtttaaa ct
//
