LOCUS       sequence-327695-        9054 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 9054)
  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 9054)
  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: AByD
FEATURES             Location/Qualifiers
     source          1..9054
                     /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(4074..4090)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4074..4090)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4087..4109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4098..4114
                     /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(4122..4128,4129..4146,4147..4152))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4167..4188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4305..4322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4476..5064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4556..4575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5235..6026,6027..6095))
                     /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     5858..5877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6096..6200)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6268..6286
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6324..6346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6446..6465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6659..6681
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6673..6690
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6674..6690
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7394..7456
                     /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 agagatctaa atgctgctga gattgaaatt taaaattggt ttatcaactg
     3001 gggcagagtg ctatatttac gggttggata acttttcccc tcctaaaatt actctctggc
     3061 ttctctggct ggctccctta cttctgttac aaagtctttc tcagtatctg gtttttatag
     3121 ttctggccgt ttttgattgg ttacaaatat aatatatctg aattttatta ttaacctagt
     3181 aatcaaatac aaaatgatta aatttaataa tataaaagaa attgatattt tccaattgta
     3241 aatttcaatg aatggattta aaataataat gttgtccaaa atatgttctg aattgttgga
     3301 ttttatataa tctcatattt ccagattcct aaagtggtta ttggttatgt agtcaagtat
     3361 tttgttttct aaattagaga ataaaatatt atttattgta gttatgcctt tgaatgttat
     3421 taaatatgaa ccgtctaaat gagaattgtt tacaatatta tttcgtatat tagtgcaaat
     3481 ttatttctct ccatttagta aacgtgaatt aaactactta agtaaattga tcatgggact
     3541 gagcaacttc atggagggaa gcgcatctca gtggcttaca caaatttcgt accatggtat
     3601 gacttagcga gagttccagg aattatttct gcagtgctat gaaaccgaag agacgccggc
     3661 cgaatgttac gcggtatatg cgtgtcgttt agtgacgacg ctgacaacga agtggcaaaa
     3721 tatgggaata gaagaaattg ccgttacaac tgttcttgcg catatggaaa gcattgacag
     3781 tcatttgcaa cacgttctct tcgttctctt ctcttcgata ttgacaccga attagtcgac
     3841 aatgagaaag ctaatatact cgagctactt aaaaagcact cgattgcatt taccaatggc
     3901 atacctgata cccgtgtaaa tacaggcgaa acgaaaatcc gtttaatcga tccaaataaa
     3961 actgtccagc gccgacttta tagacttagc cccgaagaga gcgaagtagt tcgtatgcgg
     4021 gtgtgcgaat tgataaggtg aaagcttgtt tgcccaagga attcgtaatc atggtcatag
     4081 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4141 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4201 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4261 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5341 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6121 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6181 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6241 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6301 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     6361 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6421 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     6481 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     6541 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6601 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     6661 cagtcacgac gttgtaaaac gacggccagt gccaagcttt gtttaaaaat ataacaaaat
     6721 tgtgatccca caaaatgaag tggggcaaaa tcaaataatt aatagtgtcc gtaaacttgt
     6781 tggtcttcaa ctttttgagg aacacgttgg acggcaaatc cgtgactata acacaagttg
     6841 atttaataat tttagccaac acgtcgggct gcgtgttttt tgccgacgcg tctgtgtaca
     6901 cgttgattaa ctggtcgatt aaactgttga aataatttaa tttttggttc ttctttaaat
     6961 ctgtgatgaa attttttaaa ataactttaa attcttcatt ggtaaaaaat gccacgtttt
     7021 gcaacttgtg agggtctaat atgaggtcaa actcagtagg agttttatcc aaaaaagaaa
     7081 acatgattac gtctgtacac gaacgcgtat taacgcagag tgcaaagtat aagagggtta
     7141 aaaaatatat tttacgcacc atatacgcat cgggttgata tcgttaatat ggatcaattt
     7201 gaacagttga ttaacgtgtc tctgctcaag tctttgatca aaacgcaaat cgacgaaaat
     7261 gtgtcggaca atatcaagtc gatgagcgaa aaactaaaaa ggctagaata cgacaatctc
     7321 acagacagcg ttgagatata cggtattcac gacagcaggc tgaataataa aaaaattaga
     7381 aactattatt taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgc
     7441 gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat
     7501 agatattaag ttttattata tttacactta catactaata ataaattcaa caaacaattt
     7561 atttatgttt atttatttat taaaaaaaaa caaaaactca aaatttcttc tataaagtaa
     7621 caaaactttt aaacattctc tcttttacaa aaataaactt attttgtact ttaaaaacag
     7681 tcatgttgta ttataaaata agtaattagc ttaacttata cataatagaa acaaattata
     7741 cttattagtc agtcagaaac aactttggca catatcaata ttatgctctc gacaaataac
     7801 ttttttgcat tttttgcacg atgcatttgc ctttcgcctt attttagagg ggcagtaagt
     7861 acagtaagta cgttttttca ttactggctc ttcagtactg tcatctgatg taccaggcac
     7921 ttcatttggc aaaatattag agatattatc gcgcaaatat ctcttcaaag taggagcttc
     7981 taaacgctta cgcataaacg atgacgtcag gctcatgtaa aggtttctca taaatttttt
     8041 gcgactttga accttttctc ccttgctact gacattatgg ctgtatataa taaaagaatt
     8101 tatgcaggca atgtttatca ttccgtacaa taatgccata ggccacctat tcgtcttcct
     8161 actgcagggg gacggatatt tagtctacca gtgggtgact tatctggagt ttttccattc
     8221 ttgaggatca tatgacaatc ttattcttat ctggtatctg ttaagtacat ccggagtagt
     8281 gtagggtgta tttgggtgta catgttgtgt gtgtagacat gtgcttaagt ataatggact
     8341 tatgtgtcac actatataac tataatcata tcaaaacaat tacaagtcta atcattacct
     8401 tgtttattta ggtgaatctt gttacaaaag attcacctaa tttttggatt aaactaattg
     8461 ggttttactt ggcttcccaa attctttgca acaaatgctg attcaataaa agtagcccca
     8521 gagaaaaaaa gaataagaat agaaccatta gagaacgcgc caactcgtgt gaaagtaaat
     8581 tttaatttca ttaaatgttt aattaatttg cgtcagtgtg atttttgagg tgccgtgttg
     8641 aaaatacaaa agtcacactt atgcaatgtt gtgtgtatgt gatagtgttt aaatttactc
     8701 tcttggtact ctgtcgttgt tgatttactt ttttataatt tactttatta taatttgata
     8761 tattatgggt gatagacgtt tttacttata cttttacaaa aaactttttt attgcttatc
     8821 cttaatttga aactgtcaca atctcgtccc catttagact gatcttcgaa atcttaatgc
     8881 actcaattca ctcaacctcg acctgaagtt tggtttagga ttttcttgaa cttccatgtt
     8941 taggataaaa gctttatact gaacttattt cacagcattc tgaaattcaa attcgctgac
     9001 tggtgtttag cttaagtttg gttatttatc gggtggctaa gtgctgttta aact
//
