LOCUS       sequence-327699-        9341 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 9341)
  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 9341)
  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: AByI
FEATURES             Location/Qualifiers
     source          1..9341
                     /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(4183..4199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4183..4199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4196..4218)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4207..4223
                     /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(4231..4237,4238..4255,4256..4261))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4276..4297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4414..4431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4585..5173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4665..4684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5344..6135,6136..6204))
                     /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     5967..5986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6205..6309)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6377..6395
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6433..6455)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6555..6574
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6768..6790
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6782..6799
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6783..6799
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7503..7565
                     /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 gtaatgaatg cggtaaattt gcgactctca ggtgatacta
     3001 cgatctgata atatccgctg ttcaaatcca acgtagaagg aatttgaatc gtttcgcttc
     3061 ttgtaaacgt tcttcaatgt ttggcacagg aaagttttct ttcttgatca acttgttcag
     3121 acgtcggtag tcaacacact gtcgatcgcc tccgttcttc tttttaatca aaactactgg
     3181 gctcgcatac tcagatgttg atttcgcaat aatatcgtgt tggaggagct cgtcaatcat
     3241 agcgtacaca acgtcacgct taggttcagg aattcggtat ggcgcttggg caactgcttg
     3301 tttatcttct acttcaattt cagcttcgac cacattagtt tttccaattc ctttaaggcc
     3361 agaggaaaag acataatctt ctagtaggtt ctgcatgcgt gtcttcatta ctgtcgatat
     3421 tagaaagaat attagtaata attcttgaag tgagatactt gctgtgttgc tcgtttgctc
     3481 taaaaattca tcattccgtg ctcaaacttc agcgtaagat ttgggttaac aaggatatcc
     3541 tggccgataa gaaaatcttg ctgtacgatt ttgccttcgg ctatatacat cagtgcttcg
     3601 gtgacagtat tattgatgat cagttcgatt atgattgttt cgtttagaat gcgtactcca
     3661 ccacaaatac catgaatttg cattgcacac ttgcatcttt tggcgtcgag ttgttgtgct
     3721 acagtctcac gaaccatgct agcttggcta cctgtgtcaa taaacgcagt atacggtttc
     3781 gattggacga tgatggtttt gtagaaacac ttatcttgat tcgttgcccc aaggcgtcgt
     3841 gtactcgctt cgggtttaac gcagttaccg ttgttggggt gtactttatt acacgtagta
     3901 catcggtgtc gtttcggtgg ttgtgggcat ttgttcgaaa aattacctgg ttcgccgcaa
     3961 ttaaagcatt ttaatttatc gtttgttgtt gcagactcgt gttgtttaga tttcggttca
     4021 tcgttcttcg atactggatc gtattttggc agttgaagcc tacctcggtt gacaaaatac
     4081 tcgtcaatga cgtcacgcat ctgtttcatc gttgcaaatt ttgtggtcac aatgctctgt
     4141 tgtagctctc ggtgctttaa cccagttgaa ttcgtaatca tggtcatagc tgtttcctgt
     4201 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4261 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4321 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4381 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4441 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4501 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4621 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4681 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5281 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5341 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5401 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5461 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5521 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5641 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5761 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6001 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6301 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6361 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6421 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6481 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6541 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     6601 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6661 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6721 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6781 ttgtaaaacg acggccagtg ccaagctttg tttaaaaata taacaaaatt gtgatcccac
     6841 aaaatgaagt ggggcaaaat caaataatta atagtgtccg taaacttgtt ggtcttcaac
     6901 tttttgagga acacgttgga cggcaaatcc gtgactataa cacaagttga tttaataatt
     6961 ttagccaaca cgtcgggctg cgtgtttttt gccgacgcgt ctgtgtacac gttgattaac
     7021 tggtcgatta aactgttgaa ataatttaat ttttggttct tctttaaatc tgtgatgaaa
     7081 ttttttaaaa taactttaaa ttcttcattg gtaaaaaatg ccacgttttg caacttgtga
     7141 gggtctaata tgaggtcaaa ctcagtagga gttttatcca aaaaagaaaa catgattacg
     7201 tctgtacacg aacgcgtatt aacgcagagt gcaaagtata agagggttaa aaaatatatt
     7261 ttacgcacca tatacgcatc gggttgatat cgttaatatg gatcaatttg aacagttgat
     7321 taacgtgtct ctgctcaagt ctttgatcaa aacgcaaatc gacgaaaatg tgtcggacaa
     7381 tatcaagtcg atgagcgaaa aactaaaaag gctagaatac gacaatctca cagacagcgt
     7441 tgagatatac ggtattcacg acagcaggct gaataataaa aaaattagaa actattattt
     7501 aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgcg taaaattgac
     7561 gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata gatattaagt
     7621 tttattatat ttacacttac atactaataa taaattcaac aaacaattta tttatgttta
     7681 tttatttatt aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac aaaactttta
     7741 aacattctct cttttacaaa aataaactta ttttgtactt taaaaacagt catgttgtat
     7801 tataaaataa gtaattagct taacttatac ataatagaaa caaattatac ttattagtca
     7861 gtcagaaaca actttggcac atatcaatat tatgctctcg acaaataact tttttgcatt
     7921 ttttgcacga tgcatttgcc tttcgcctta ttttagaggg gcagtaagta cagtaagtac
     7981 gttttttcat tactggctct tcagtactgt catctgatgt accaggcact tcatttggca
     8041 aaatattaga gatattatcg cgcaaatatc tcttcaaagt aggagcttct aaacgcttac
     8101 gcataaacga tgacgtcagg ctcatgtaaa ggtttctcat aaattttttg cgactttgaa
     8161 ccttttctcc cttgctactg acattatggc tgtatataat aaaagaattt atgcaggcaa
     8221 tgtttatcat tccgtacaat aatgccatag gccacctatt cgtcttccta ctgcagggca
     8281 tacaggggaa ccagaattga tttatacttc ggctgattgt tttgtcggtg acattctaca
     8341 agaatgcgat agagtaactg ctttggtaca acccaaaggt gcttgttgtt atgtttgcga
     8401 tataagcgac caagttcaag gacatactca tcgttctcag tgatcttctt agcaatcatt
     8461 tcgctagcaa ttcgttttat tttctcgtcc tgcaattgta cagcaaacaa ccaatcggct
     8521 tcatcaatga cggttttgaa tatctttagg tcagcttctt ccatttcacg cgaatcttca
     8581 tatggtgatc ggcttaaggc atcgacgtgt tccatacacg taactgagcg atgcagtatc
     8641 tcaatatcga attcttggat gcgtagccac catctggcaa ttcggggtat cagctctcgt
     8701 ttctccatcg tcgtttttaa cgcattacaa tcagtgatga ctcgaattcg tttcccatag
     8761 acataatact tgaaacgttc cagtgactcg acaacagcca aagcttctag ctcgtagatg
     8821 tgaaacttca tttcgctgta tgtcgtcgat cggctataat aggctactgg ctttaactga
     8881 tcatgttcct ttgcaataga actccggcta aacaaacaga actagcatca gtatgtattt
     8941 cgtgttctgg gctgacgtcg tagctaataa gaacaggctt cgaaataagg agttgtttaa
     9001 gtttattgaa tgcgtcatcc tgctgtggtt cccatttaaa cggctggtct ttgcgtagta
     9061 gcttgcgtaa ggcctcggaa ataatggcgt atcctgggat aaatttttcg tacatccgaa
     9121 acgttcttta gacgtggaag ctatgcaatg gcattggttt tgacttcacc tggacgtatc
     9181 ccggttgcgt gcagctggtg ccccaagaat gaaattgtct gcttgtacaa ttcgcacttg
     9241 tcaacgttca gggtgaggcc acattctcgt agcacttgca gtatctcggt aaacgaatgg
     9301 ctgccaatta atgtcgtcca tatctcccgt tcgtttaaac t
//
