LOCUS       sequence-327688-        8798 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 8798)
  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 8798)
  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: AByB
FEATURES             Location/Qualifiers
     source          1..8798
                     /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(3798..3814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3798..3814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3811..3833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3822..3838
                     /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(3846..3852,3853..3870,3871..3876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3891..3912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4029..4046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4200..4788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4280..4299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4959..5750,5751..5819))
                     /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     5582..5601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5820..5924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5992..6010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6048..6070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6170..6189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6383..6405
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6397..6414
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6398..6414
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7118..7180
                     /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 agagatcttt aaaactctgt tttttgtgag catttcaggt ttgaggtcgc
     3001 aaattgctct aatctacttt attttagttt tatatgataa aatatagtta aaatatataa
     3061 gaatttacgt ttaattaata tttaaacgat ttcttactgc taaatgggta gttctttgaa
     3121 tactctttga caacttgtat aattttttat attcgagctc tttagttgtt aatttctttt
     3181 tataatctcg caagaataaa aatctctgca agaataaaaa caaaacaaca gccaaacaag
     3241 cggctttcgt tgacaatttg gacaaccttt tctatattgc acatgcaaat gcaatggata
     3301 tgatgtcaat tgaagaggat aagaaattcc atgaaattcc attttaacac ccagagttat
     3361 ttcattgttg gacaaatgac agatttccaa taatgcagct atgcatttga cttctgcggt
     3421 tgttgaagct ttgggtcatg atatcgagga attcgcattc cataccagaa cttttcagca
     3481 aagctgtaaa gacttggtaa ttaccttgca actctttgaa tgttcccatt ttaacatgta
     3541 atttcagatg actgcagtgg tacattggga tggaaaaacg ctcagaattg attggtcgag
     3601 aaaaagtcga aagaatacca attataatat catattctga tggtgaaaaa cttttgggag
     3661 taccaaaagt agttgcatcc actggcgttg aaatatctca tgcaaccttc aacatgctaa
     3721 cagagtgggg aatttcggat aaaatcgttg ctgtatgttt tgaaacaact gcatcgaatt
     3781 ccgaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3841 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3901 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3961 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4021 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4081 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4141 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4381 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4621 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4861 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4921 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4981 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5041 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5101 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5161 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5221 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5281 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5341 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5401 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5461 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5521 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5581 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5641 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5701 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5761 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5821 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5881 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5941 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6001 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6061 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6121 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     6181 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     6241 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6301 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     6361 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     6421 ctttgtttaa aaatataaca aaattgtgat cccacaaaat gaagtggggc aaaatcaaat
     6481 aattaatagt gtccgtaaac ttgttggtct tcaacttttt gaggaacacg ttggacggca
     6541 aatccgtgac tataacacaa gttgatttaa taattttagc caacacgtcg ggctgcgtgt
     6601 tttttgccga cgcgtctgtg tacacgttga ttaactggtc gattaaactg ttgaaataat
     6661 ttaatttttg gttcttcttt aaatctgtga tgaaattttt taaaataact ttaaattctt
     6721 cattggtaaa aaatgccacg ttttgcaact tgtgagggtc taatatgagg tcaaactcag
     6781 taggagtttt atccaaaaaa gaaaacatga ttacgtctgt acacgaacgc gtattaacgc
     6841 agagtgcaaa gtataagagg gttaaaaaat atattttacg caccatatac gcatcgggtt
     6901 gatatcgtta atatggatca atttgaacag ttgattaacg tgtctctgct caagtctttg
     6961 atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca agtcgatgag cgaaaaacta
     7021 aaaaggctag aatacgacaa tctcacagac agcgttgaga tatacggtat tcacgacagc
     7081 aggctgaata ataaaaaaat tagaaactat tatttaaccc tagaaagata atcatattgt
     7141 gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg gtctgtatat
     7201 cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca cttacatact
     7261 aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa aaaacaaaaa
     7321 ctcaaaattt cttctataaa gtaacaaaac ttttaaacat tctctctttt acaaaaataa
     7381 acttattttg tactttaaaa acagtcatgt tgtattataa aataagtaat tagcttaact
     7441 tatacataat agaaacaaat tatacttatt agtcagtcag aaacaacttt ggcacatatc
     7501 aatattatgc tctcgacaaa taactttttt gcattttttg cacgatgcat ttgcctttcg
     7561 ccttatttta gaggggcagt aagtacagta agtacgtttt ttcattactg gctcttcagt
     7621 actgtcatct gatgtaccag gcacttcatt tggcaaaata ttagagatat tatcgcgcaa
     7681 atatctcttc aaagtaggag cttctaaacg cttacgcata aacgatgacg tcaggctcat
     7741 gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt tctcccttgc tactgacatt
     7801 atggctgtat ataataaaag aatttatgca ggcaatgttt atcattccgt acaataatgc
     7861 cataggccac ctattcgtct tcctactgca ggtttaaatg ggtttgccct aatttttaaa
     7921 aggttgaaat gtcatgcccc gctaaggtca tgaagaacct atcgaggagt gagtagaatt
     7981 gagttttttt tttatatatt aattcggtca ttttcttatg ctaacttata tgttattaca
     8041 gttgacatgt cttctaattg tttacagaat agacgtaagt tatcttgaaa aggtgtgcga
     8101 tataaccttc ctagtagttc ctctagaggc atctgactct tgaatttcgt gattagggcg
     8161 tcttttaaat ttgtccattc gttgtgcatt ccaaattggg aagcctttcc gcatctccga
     8221 caatttgaag ttcggtggcg tcaaatatta ttgccttttg tcgaatgtcg ttcgaatggt
     8281 ataagttcga tacgtctgat gaacgagatg agaatccata gaattacgat acttggcgta
     8341 tttgcctaag taactgagac agatttttcg ctgttaacgc cattttagtt ttatttttag
     8401 attagtttcc gttcttgttc acagactggt aacgaatgac gagacttatg tcttagtgaa
     8461 ctgcactttc tccttattac accttagtgg cggtctcact tcccgaagca ggccacaaaa
     8521 ttaggtttta aaattaacta ggatctcaca gtttttgtgc gggtaccgat tttatatagt
     8581 ttaaacgaac cgaaatcact ttgttgcggc caattcgcgg gtacaatcaa aaaagcttac
     8641 ttggtcatta gggttcggct tattggtatg gagaaaaaat tgcgaaaatt aaaggttcaa
     8701 ggtctgaaat tgttggtttt ggtgtaaaac ttaaatttgt aaagttttag cccgatcgga
     8761 taatatttag aggtgccttt cttaatatag tttaaact
//
