LOCUS       sequence-208911-        8687 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 8687)
  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 8687)
  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..8687
                     /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    4593..5023
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     regulatory      5292..5301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5431..5449)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5699..5718
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5893..5916
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    6850..6971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6887..6906)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6941..6960
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    7049..7148
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(8380..8396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8380..8396)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8393..8415)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8404..8420
                     /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(8428..8434,8435..8452,8453..8458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8473..8494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8611..8628)
                     /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 ggtgtacctc
     3781 gcccagttgg tcctattggt tgagggccta cacggtcgct cgatgtgttg tagttggtgc
     3841 aaaagatgaa agattaccgg acagtgacca gatgagagtt ctggaagtgc ctcagctgta
     3901 atcattgatt gggatatcct ttttgtgatg gcaaagtcaa tcaaatctgg aagcttcatg
     3961 ggatctgctg gccattatgt aggcgctccc ggggaaacat agttaagttt gtgttgcggt
     4021 ttgatgattg cgttgtacag caacaaacct ttcaccaaac gagtgaaaag tgttgtgaat
     4081 ttttttctcc aacgccaacg cttaactata ctctataact actcattcac caaaatctat
     4141 ttgctctatc cataaactag ttctttccaa gtaacttaat ctcccgaatc gtaatttctc
     4201 actcgaacta tggcgcgcat agttgtgctt aaggttggag gtgaggtgtt ttgtagatgc
     4261 ccgaagtcca cctgtggctt gttcggtgac aaaaactgat ctgattctat attctagctc
     4321 tggtctcaat tagacctccg ctcagagaag taaataggaa atagtgtctt aaatgctaca
     4381 tttgtaagtg ttaattcata agtgctgcgt cgcacttatt ccgaattgct gtgttgccta
     4441 ctcttgagtg gatcgattct ttgctgagtc gttctcctat tacatagtcc ggaggagtag
     4501 gggtttcctg tccaaatgtc atgttaacgg ccgttcgctg aacggtcgtt gtgtggccta
     4561 attaaatccg ttgtttacat tttgtttaaa cttggccacg taataagtgt gcgttgaatt
     4621 tattcgcaaa aacattgcat attttcggca aagtaaaatt ttgttgcata ccttatcaaa
     4681 aaataagtgc tgcatacttt ttagagaaac caaataattt tttattgcat acccgttttt
     4741 aataaaatac attgcatacc ctcttttaat aaaaaatatt gcatactttg acgaaacaaa
     4801 ttttcgttgc atacccaata aaagattatt atattgcata cccgttttta ataaaataca
     4861 ttgcataccc tcttttaata aaaaatattg catacgttga cgaaacaaat tttcgttgca
     4921 tacccaataa aagattatta tattgcatac cttttcttgc cataccattt agccgatcaa
     4981 ttgtgctcgg caacagtata tttgtggtgt gccaaccaac aacggcgcgc ctaaattcga
     5041 gctcgcccgg ggatctaatt caattagaga ctaattcaat tagagctaat tcaattagga
     5101 tccaagctta tcgatttcga accctcgacc gccggagtat aaatagaggc gcttcgtcta
     5161 cggagcgaca attcaattca aacaagcaaa gtgaacacgt cgctaagcga aagctaagca
     5221 aataaacaag cgcagctgaa caagctaaac aatcggggta ccgctagagt cgacggtacg
     5281 atccaccggt cgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct
     5341 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     5401 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     5461 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
     5521 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     5581 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     5641 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     5701 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     5761 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     5821 tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     5881 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     5941 acgagcgctc cgagggccgc caccacctgt tcctggggca tggcaccggc agcaccggca
     6001 gcggcagctc cggcaccgcc tcctccgagg acaacaacat ggccgttatc aaggaattta
     6061 tgcgcttcaa agttaggatg gagggatcca tgaacggaca tgagttcgag atcgagggag
     6121 agggcgaggg acgcccgtat gaaggcacac aaacagccaa actcaaggtc accaagggcg
     6181 gaccactgcc cttcgcctgg gatatcctga gtccccagtt tatgtacggc agcaaggcct
     6241 acgttaagca ccccgctgac ataccggact acaaaaagct gtcctttccg gaaggcttca
     6301 agtgggagcg cgtgatgaat ttcgaagacg gaggactggt cactgtgacc caagatagca
     6361 gtttgcagga cggtacactg atctataagg ttaaaatgcg cggcactaac tttccgccag
     6421 atggcccagt gatgcagaag aagaccatgg gttgggaggc atccaccgaa cgtctgtacc
     6481 ctcgagacgg agtgctcaag ggcgagatcc atcaggccct caaactgaaa gatggtggtc
     6541 actacctggt cgaatttaag accatttaca tggccaagaa gccggttcag ctgcccggat
     6601 attattatgt ggatacgaaa ctggatataa cttcgcataa cgaagactac accattgtcg
     6661 agcagtatga gcgcagcgaa ggccgacatc acctgttcct ctacggcatg gacgagctgt
     6721 acaagtaggc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     6781 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     6841 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6961 aatgtatctt aaagcttcct agggtactga cggacacacc gaagccccgg cggcaaccct
     7021 cagcggatgc cccggggctt cacgttttcc caggtcagaa gcggttttcg ggagtagtgc
     7081 cccaactggg gtaacctttg agttctctca gttgggggcg tagggccgcc gacatgacac
     7141 aaggggttgt gaccggggtg gacacgtacg cgggtgctta cgaccgtcag tcgcgcgagc
     7201 gcgagggccc gggtttttta aatggtaccc cttgcagcgc cacctggccg cgaagagttc
     7261 cattctctgc ttcgaagtac gaagagagaa agagtacttt aaatttccac attcccgcct
     7321 tgcagcgcca cctggccttg gtaatgtaga actaggaagg aaagcaccaa cacaagatgt
     7381 cgctctccga cgatggacat gtcgcgtaaa aaatgttcga taactttcaa tggttcgatt
     7441 gaacagacaa taagtgtatt taagacacca gttcttatat tcaaaaatcc taacaactca
     7501 caccttgcag cgccacctgg ccctagagat ccaccaactc tagagagatc tgtaagccgg
     7561 cgtggcttag ctagagggta tgtgtgccgt ttggaaccgg taactcctcc ctccttggat
     7621 tgaagtgaca gccacaacga actagtagtc ttcactaagc tcggctctgg taccgtttta
     7681 aatgccactc ttcttcttct attgatgagg tatattgtcg tcactactcc aactaatagt
     7741 aatacgtaaa gtgctccgaa atttgtagtc gtaaattttt tccaatatat gatggaatta
     7801 gaagcatggt tggatcttat gttcagctta tgcaggttaa tgtcttctgt gttctcggtg
     7861 aaatatatat tttttataat tggggattgt gcggatcttc ttatggaaat tttggtacaa
     7921 tttgattcaa cggtagtgtt ttgtgtgttt aatattaaaa tgtgagctgg ttctacttct
     7981 tttataattt cttgattccc tatgtcttta gttttaaaat tggcagtttc catcttatgt
     8041 atgttattga gacattcagt tttaatatta cctctgattg ccattgcatt gtcgcagata
     8101 tggtatttat atatcttgtt atatttgttg gaaaagtagt aaaaaacatg ccatatagtt
     8161 aggtatattt atatatttat atttatattt catttatggg taatgggaca atgtgtgtga
     8221 acttttagta attttgtttg aatgcaggga ttcttattct aaagtttatc ttaattccaa
     8281 tcatgaaagt atctaattct aacatttgaa atatgtgttt gtctgacatt tcccgtaact
     8341 ccgggtcgtt taattcttgt ctgagaattc gtaatcatgg tcatagctgt ttcctgtgtg
     8401 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     8461 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     8521 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     8581 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     8641 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggt
//
