LOCUS       sequence-327685-        8747 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 8747)
  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 8747)
  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: AByA
FEATURES             Location/Qualifiers
     source          1..8747
                     /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(3675..3691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3675..3691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3688..3710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3699..3715
                     /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(3723..3729,3730..3747,3748..3753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3768..3789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3906..3923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4077..4665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4157..4176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4836..5627,5628..5696))
                     /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     5459..5478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5697..5801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5869..5887
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5925..5947)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6047..6066
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6260..6282
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6274..6291
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6275..6291
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6995..7057
                     /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 gaggagcaga tcaacaagga atcaaaagaa atgacatcaa
     3001 gaccaattgc ggaacccatc tgtaaaatta cacatcgtgg ttttaaacag agtggggagt
     3061 gccccagatg tgggaaattc ggacacacac acaacgaccc atcatgtccg gcaagaaatg
     3121 tgacttgcaa taaatgctcg aagcctggac actttgccag gaagtgccga actaacttga
     3181 acggccgatt ccttaagtcc caaaggaata acctaaaaag gcctcgcacg tatattcgct
     3241 ctgtcgaaga ggaggccacc agctccaaac tcaaaaaagg ggaggcacat tgctttaaag
     3301 tgtcaagtgt agatgaagaa gagttcatac gatgccgagt gggaggccgt gaaatgtctt
     3361 taattattga ctcggggtgc aagtttaacc ttatcaccca cccggactgg tctatgctag
     3421 tgaagagaaa agccacagtc tttaccgtaa gaacccatat agaaaagcaa ttccgagcct
     3481 atgcctctaa taaattacta cgagtaatgt atattttcga ggccccaatt tccgttgagc
     3541 cgggtaccga agagattgcc tcattttacg tgattgaaaa tggagaacaa accagactca
     3601 aagaggctaa attcttcgca cgcctagacc tcaaggacgc ctatcaccaa ctggcccttg
     3661 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5821 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5881 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5941 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6001 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6061 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6121 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6181 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6241 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
     6301 tgtttaaaaa tataacaaaa ttgtgatccc acaaaatgaa gtggggcaaa atcaaataat
     6361 taatagtgtc cgtaaacttg ttggtcttca actttttgag gaacacgttg gacggcaaat
     6421 ccgtgactat aacacaagtt gatttaataa ttttagccaa cacgtcgggc tgcgtgtttt
     6481 ttgccgacgc gtctgtgtac acgttgatta actggtcgat taaactgttg aaataattta
     6541 atttttggtt cttctttaaa tctgtgatga aattttttaa aataacttta aattcttcat
     6601 tggtaaaaaa tgccacgttt tgcaacttgt gagggtctaa tatgaggtca aactcagtag
     6661 gagttttatc caaaaaagaa aacatgatta cgtctgtaca cgaacgcgta ttaacgcaga
     6721 gtgcaaagta taagagggtt aaaaaatata ttttacgcac catatacgca tcgggttgat
     6781 atcgttaata tggatcaatt tgaacagttg attaacgtgt ctctgctcaa gtctttgatc
     6841 aaaacgcaaa tcgacgaaaa tgtgtcggac aatatcaagt cgatgagcga aaaactaaaa
     6901 aggctagaat acgacaatct cacagacagc gttgagatat acggtattca cgacagcagg
     6961 ctgaataata aaaaaattag aaactattat ttaaccctag aaagataatc atattgtgac
     7021 gtacgttaaa gataatcatg cgtaaaattg acgcatgtgt tttatcggtc tgtatatcga
     7081 ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt acatactaat
     7141 aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa acaaaaactc
     7201 aaaatttctt ctataaagta acaaaacttt taaacattct ctcttttaca aaaataaact
     7261 tattttgtac tttaaaaaca gtcatgttgt attataaaat aagtaattag cttaacttat
     7321 acataataga aacaaattat acttattagt cagtcagaaa caactttggc acatatcaat
     7381 attatgctct cgacaaataa cttttttgca ttttttgcac gatgcatttg cctttcgcct
     7441 tattttagag gggcagtaag tacagtaagt acgttttttc attactggct cttcagtact
     7501 gtcatctgat gtaccaggca cttcatttgg caaaatatta gagatattat cgcgcaaata
     7561 tctcttcaaa gtaggagctt ctaaacgctt acgcataaac gatgacgtca ggctcatgta
     7621 aaggtttctc ataaattttt tgcgactttg aaccttttct cccttgctac tgacattatg
     7681 gctgtatata ataaaagaat ttatgcaggc aatgtttatc attccgtaca ataatgccat
     7741 aggccaccta ttcgtcttcc tactgcagga gcaaccaaag agttgcaagg gccgagtacg
     7801 cattataagc tctaaaagag gagccgagta gtggtacaag ccaaagcata ttttgaaaaa
     7861 aaagaaaaga aacagaacac gaaatatata aataaataaa tattttaata attaaaaatt
     7921 tttgcttaat taaaaaaaaa aacagaagga aaaaggaaaa atattacaag cttttaagaa
     7981 agaactgggt aatcggtttg agttgcaaac aaaaggttaa aggtcgggtg gagcagtatt
     8041 ctctccacaa gccaaaagaa ttcataaaat ttaaaagaaa aaacggaaaa agcaaaaccc
     8101 caaatgtaaa tgggaggcat tattaaactg aggctgggta ataattacaa gctcctaaga
     8161 gagccgggta ttaattacaa gtcaagtcgt aaatctaata atgtctttcg tcgtaaactc
     8221 caattcgagg tcccgtattg aaaatgaccg acagtgtcat taaaccgttt ctgggcgagg
     8281 tgatcgacaa ggcaatcctt cgaaatgaat gggagaagtg gttgagggcg tttacaatat
     8341 acctcgaggc cgagggtatt gataccgaga agcagaagag gagtaagcta ctgcttttgg
     8401 gaggagtcca gctacagtca gtagtctacc tggtgcgctg gtggagccca gcgacggaaa
     8461 taaagaggac atctataaaa tcctcatcga ccatttaaat aaacacttct caccgaagca
     8521 gaactcaacg tttgaaagac atctattcag aggcctgaca ccgttggaca cagaaagctt
     8581 tggtgatttc atgctgcgac tccgccaaca aatgcaatga tgtgcattcg gatcctcaaa
     8641 agcggagatt gagaatatat gtctaaagga taaaatcata gatgtgtggg cacctctaga
     8701 cctcaagaaa agactcctgg gaaaggagta ttcactaggt ttaaact
//
