LOCUS       sequence-318530-        8963 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 8963)
  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 8963)
  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: AByC
FEATURES             Location/Qualifiers
     source          1..8963
                     /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   2998..3060
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4641..5071
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     regulatory      5340..5349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5479..5497)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5747..5766
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5941..5964
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    6898..7019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6935..6954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6989..7008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    7097..7196
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(8656..8672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8656..8672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8669..8691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8680..8696
                     /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(8704..8710,8711..8728,8729..8734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8749..8770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8887..8904)
                     /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 gctttctcaa tgctcacgct
      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 agaaggacag
      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 tttaaaatat aacaaaattg
     2341 tgatcccaca aaatgaagtg gggcaaaatc aaataattaa ctagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcg tgactataac acaagttgat
     2461 ttaataattt tagccaacac gtcgggctgc gtgttttttg cgctctgtgt acacgttgat
     2521 taactggtcg attaaataat ttaatttttg gttcttcttt aaatctgtga tgaaattttt
     2581 taaaataact ttaaattctt cattggtaaa aaatgccacg ttttgcaact tgtgagggtc
     2641 taatatgagg tcaaactcag taggagtttt atccaaaaaa gaaaacatga ttacgtctgt
     2701 acacgaacgc gtattaacgc agagtgcaaa gtataagagg gttaaaaaat atattttacg
     2761 caccatatac gcatcgggtt gatatcgtta atatggatca atttgaacag ttgattaacg
     2821 tgtctctgct caagtctttg atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca
     2881 agtcgatgag cgaaaaacta aaaaggctag aatacgacaa tctcacagac agcgttgaga
     2941 tatacggtat tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc
     3001 tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg
     3061 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
     3121 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
     3181 ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat
     3241 tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa
     3301 aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt agtcagtcag
     3361 aaacaacttt ggcacatatc aatattatgc tctcgacaaa taactttttt gcattttttg
     3421 cacgatgcat ttgcctttcg ccttatttta gaggggcagt aagtacagta agtacgtttt
     3481 ttcattactg gctcttcagt actgtcatct gatgtaccag gcacttcatt tggcaaaata
     3541 ttagagatat tatcgcgcaa atatctcttc aaagtaggag cttctaaacg cttacgcata
     3601 aacgatgacg tcaggctcat gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt
     3661 tctcccttgc tactgacatt atggctgtat ataataaaag aatttatgca ggcaatgttt
     3721 atcattccgt acaataatgc cataggccac ctattcgtct tcctactgca gggttgtata
     3781 aatttatttt ctgatcaaat tgatgttgtc ttttcggagg agtttgttta agaggagttt
     3841 gattgtttac gttagtgttt tgggtctttt aatggatttg ttaatggaat aaaggggcga
     3901 atgaatcgtt ggttgcggtg agtgtttttg ctgactatcg gtttttcttt tttttaaatg
     3961 ggtttgccct aatttttaaa aggttgaaat gtcatgcccc gctaaggtca tgaagagcct
     4021 atcgaggagt gagtagaatt gagttttttt tttatagtag ttgccattgc ttgtgtataa
     4081 accggtatat tatttcggtc attttcttat gctaacttat atgttattac agttgacatg
     4141 tcttctaatt gtttacagaa tagacgtaag ttatcttgaa aaggtgtgcg atataacctt
     4201 cctagtagtt cctctagagg catctgactc ttgaatttcg tgattagggc gtcttttaaa
     4261 tttgtccatt cgttgtgcat tccaaattgg gaagcctttc cgcatctccg acaatttgaa
     4321 gttcggtggc gtcaaatatt attgcctttt gtcgtatgtc gttcgaatgg tataagttcg
     4381 atacgtctga tgaacgagat gagaatccat agaattacga tacttggcgt atttgcctaa
     4441 gtaactgaga cagatttttc gctgttaacg ccattttagt tttattttta gattagtttc
     4501 cgttcttgtt cacagactgg taacgaatga cgagacttat gtctcagtga actgcacttt
     4561 ctccttatta caccttagtg gcggtctcac ttcccgaagc aggccacaaa attaggtttt
     4621 aaaattaact agtttaaact tggccacgta ataagtgtgc gttgaattta ttcgcaaaaa
     4681 cattgcatat tttcggcaaa gtaaaatttt gttgcatacc ttatcaaaaa ataagtgctg
     4741 catacttttt agagaaacca aataattttt tattgcatac ccgtttttaa taaaatacat
     4801 tgcataccct cttttaataa aaaatattgc atactttgac gaaacaaatt ttcgttgcat
     4861 acccaataaa agattattat attgcatacc cgtttttaat aaaatacatt gcataccctc
     4921 ttttaataaa aaatattgca tacgttgacg aaacaaattt tcgttgcata cccaataaaa
     4981 gattattata ttgcatacct tttcttgcca taccatttag ccgatcaatt gtgctcggca
     5041 acagtatatt tgtggtgtgc caaccaacaa cggcgcgcct aaattcgagc tcgcccgggg
     5101 atctaattca attagagact aattcaatta gagctaattc aattaggatc caagcttatc
     5161 gatttcgaac cctcgaccgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat
     5221 tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg
     5281 cagctgaaca agctaaacaa tcggggtacc gctagagtcg acggtacgat ccaccggtcg
     5341 ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca
     5401 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     5461 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     5521 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     5581 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     5641 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     5701 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     5761 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     5821 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     5881 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     5941 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     6001 agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc ggcagctccg
     6061 gcaccgcctc ctccgaggac aacaacatgg ccgttatcaa ggaatttatg cgcttcaaag
     6121 ttaggatgga gggatccatg aacggacatg agttcgagat cgagggagag ggcgagggac
     6181 gcccgtatga aggcacacaa acagccaaac tcaaggtcac caagggcgga ccactgccct
     6241 tcgcctggga tatcctgagt ccccagttta tgtacggcag caaggcctac gttaagcacc
     6301 ccgctgacat accggactac aaaaagctgt cctttccgga aggcttcaag tgggagcgcg
     6361 tgatgaattt cgaagacgga ggactggtca ctgtgaccca agatagcagt ttgcaggacg
     6421 gtacactgat ctataaggtt aaaatgcgcg gcactaactt tccgccagat ggcccagtga
     6481 tgcagaagaa gaccatgggt tgggaggcat ccaccgaacg tctgtaccct cgagacggag
     6541 tgctcaaggg cgagatccat caggccctca aactgaaaga tggtggtcac tacctggtcg
     6601 aatttaagac catttacatg gccaagaagc cggttcagct gcccggatat tattatgtgg
     6661 atacgaaact ggatataact tcgcataacg aagactacac cattgtcgag cagtatgagc
     6721 gcagcgaagg ccgacatcac ctgttcctct acggcatgga cgagctgtac aagtaggcgg
     6781 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     6841 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     6901 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6961 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     7021 agcttcctag ggtactgacg gacacaccga agccccggcg gcaaccctca gcggatgccc
     7081 cggggcttca cgttttccca ggtcagaagc ggttttcggg agtagtgccc caactggggt
     7141 aacctttgag ttctctcagt tgggggcgta gggccgccga catgacacaa ggggttgtga
     7201 ccggggtgga cacgtacgcg ggtgcttacg accgtcagtc gcgcgagcgc gagggcccgg
     7261 gttttttaaa tggtacccct tgcagcgcca cctggccgcg aagagttcca ttctctgctt
     7321 cgaagtacga agagagaaag agtactttaa atttccacat tcccgccttg cagcgccacc
     7381 tggccttggt aatgtagaac taggaaggaa agcaccaaca caagatgtcg ctctccgacg
     7441 atggacatgt cgcgtaaaaa atgttcgata actttcaatg gttcgattga acagacaata
     7501 agtgtattta agacaccagt tcttatattc aaaaatccta acaactcaca ccttgcagcg
     7561 ccacctggcc ctagagatcc accaactcta gagagatctt accgatttta tatagtttaa
     7621 acgaaccgaa atcactttgt tgcggccaat tcgcgggtac aatcagaaaa gcttacttgg
     7681 tcatgtcgcc acatgtataa acaaagaaaa taatgagatc attgatttct gtcgatgtca
     7741 attgaaaaaa gattacgaac gttattacta ccgcgaacta ttagagttaa caattgtctt
     7801 tttgggaata tttccggaag gccatgattt ccgaccgccc ggccctatgc atcatgctcg
     7861 ctggatgtct aaggcaatta atgcactgaa gctatacctt tttcgagaag tcttccgttt
     7921 aactaaacac gaagacaagt agttgcgtga tgtgtctgtt tttatagtga caatttatgt
     7981 aaaattgttg tttggatccc tgttggtcct tgaagctcct tatagagaat ttcaatttat
     8041 tcgggctgtg tacatctaca gagcagtgga cgagaggatt tcaatggcag tgctcaaaaa
     8101 gacgtagtca tttatggtac ctggcagacg aaaatatcac attggctcta tttgatgata
     8161 gcgttactgt cgcagaaaat aagaaatggc acagcgctta ttaaccgttg cggatgatac
     8221 aggcaaagat gaacattccg ttaagcgcat tgatttaaaa gtatgagatt ttccaacttt
     8281 tgtttcaaaa aaattatctg atttcattac atcaaatagt attaagtttt tcgaccggtt
     8341 caatgtatcg aaatcttctt tagtcataga tccatcgaag ttgcatgatc acgaagactt
     8401 taaaaatgct gttgagattg tacgaaaatt aaaatctgat tataactaga aattaacaac
     8461 taaagacctc gaatatcaat aattattaca aattgtcaat gagtatacaa agaagtacac
     8521 atttagcagt aaaaaatccc ttaaaaaaca ttactaaaca ttcttttctt tttctctttt
     8581 cactatattg tcaaaggata acgagcttac taggtttttc agtatcctac cgactgcacc
     8641 gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8701 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8761 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8821 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8941 tcactcaaag gcggtaatac ggt
//
