LOCUS       sequence-170967-        7213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE Bxb1 attB-flanked vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7213)
  AUTHORS   Voutev R, Mann RS
  TITLE     Bxb1 phage recombinase assists genome engineering in Drosophila 
            melanogaster.
  JOURNAL   Biotechniques. 2017 Jan 1;62(1):37-38. doi: 10.2144/000114494.
  PUBMED    28118814
REFERENCE   2  (bases 1 to 7213)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2017-01-1- 1"; volume: "62"; issue: "1"; 
            pages: "37-38"
COMMENT     Sequence Label: pRVV601
FEATURES             Location/Qualifiers
     source          1..7213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            665..4801
                     /gene="white"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
     primer_bind     complement(674..692)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1144..1215,1617..1890,1965..2619,2681..2996,3217..3348,
                     3419..4033)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4835..4872
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    4963..5000
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        complement(5025..5043)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5064..5080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5064..5080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5077..5099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5088..5104
                     /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(5112..5118,5119..5136,5137..5142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5157..5178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5295..5312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5466..6054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5546..5565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6225..7016,7017..7085))
                     /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     6848..6867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7086..7190)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggccctagta tgtatgtaag ttaataaaac ccatttttgc ggaaagtaga taaaaaaaac
      721 attttttttt tttactgcac tggatatcat tgaacttatc tgatcagttt taaatttact
      781 tcgatccaag ggtatttgat gtaccaggtt ctttcgatta cctctcactc aaaatgacat
      841 tccactcaaa gtcagcgctg tttgcctcct tctctgtcca cagaaatatc gccgtctctt
      901 tcgccgctgc gtccgctatc tctttcgcca ccgtttgtag cgttacgtag cgtcaatgtc
      961 cgccttcagt tgcattttgt cagcggtttc gtgacgaagc tccaagcggt ttacgccatc
     1021 aattaaacac aaagtgctgt gccaaaactc ctctcgcttc ttatttttgt ttgttttttg
     1081 agtgattggg gtggtgattg gttttgggtg ggtaagcagg ggaaagtgtg aaaaatcccg
     1141 gcaatgggcc aagaggatca ggagctatta attcgcggag gcagcaaaca cccatctgcc
     1201 gagcatctga acaatgtgag tagtacatgt gcatacatct taagttcact tgatctatag
     1261 gaactgcgat tgcaacatca aattgtctgc ggcgtgagaa ctgcgaccca caaaaatccc
     1321 aaaccgcaat tgcacaaaca aatagtgaca cgaaacagat tattctggta gctgttctcg
     1381 ctatataaga caatttttga gatcatatca tgatcaagac atctaaaggc attcattttc
     1441 gactatattc ttttttacaa aaaatataac aaccagatat tttaagctga tcctagatgc
     1501 acaaaaaata aataaaagta taaacctact tcgtaggata cttcggggta ctttttgttc
     1561 ggggttagat gagcataacg cttgtagttg atatttgaga tcccctatca ttgcagggtg
     1621 acagcggagc ggcttcgcag agctgcatta accagggctt cgggcaggcc aaaaactacg
     1681 gcacgctccg gccacccagt ccgccggagg actccggttc agggagcggc caactagccg
     1741 agaacctcac ctatgcctgg cacaatatgg acatctttgg ggcggtcaat cagccgggct
     1801 ccggatggcg gcagctggtc aaccggacac gcggactatt ctgcaacgag cgacacatac
     1861 cggcgcccag gaaacatttg ctcaagaacg gtgagtttct attcgcagtc ggctgatctg
     1921 tgtgaaatct taataaaggg tccaattacc aatttgaaac tcagtttgcg gcgtggccta
     1981 tccgggcgaa cttttggccg tgatgggcag ttccggtgcc ggaaagacga ccctgctgaa
     2041 tgcccttgcc tttcgatcgc cgcagggcat ccaagtatcg ccatccggga tgcgactgct
     2101 caatggccaa cctgtggacg ccaaggagat gcaggccagg tgcgcctatg tccagcagga
     2161 tgacctcttt atcggctccc taacggccag ggaacacctg attttccagg ccatggtgcg
     2221 gatgccacga catctgacct atcggcagcg agtggcccgc gtggatcagg tgatccagga
     2281 gctttcgctc agcaaatgtc agcacacgat catcggtgtg cccggcaggg tgaaaggtct
     2341 gtccggcgga gaaaggaagc gtctggcatt cgcctccgag gcactaaccg atccgccgct
     2401 tctgatctgc gatgagccca cctccggact ggactcattt accgcccaca gcgtcgtcca
     2461 ggtgctgaag aagctgtcgc agaagggcaa gaccgtcatc ctgaccattc atcagccgtc
     2521 ttccgagctg tttgagctct ttgacaagat ccttctgatg gccgagggca gggtagcttt
     2581 cttgggcact cccagcgaag ccgtcgactt cttttcctag tgagttcgat gtgtttatta
     2641 agggtatcta gcattacatt acatctcaac tcctatccag cgtgggtgcc cagtgtccta
     2701 ccaactacaa tccggcggac ttttacgtac aggtgttggc cgttgtgccc ggacgggaga
     2761 tcgagtcccg tgatcggatc gccaagatat gcgacaattt tgctattagc aaagtagccc
     2821 gggatatgga gcagttgttg gccaccaaaa atttggagaa gccactggag cagccggaga
     2881 atgggtacac ctacaaggcc acctggttca tgcagttccg ggcggtcctg tggcgatcct
     2941 ggctgtcggt gctcaaggaa ccactcctcg taaaagtgcg acttattcag acaacggtga
     3001 gtggttccag tggaaacaaa tgatataacg cttacaattc ttggaaacaa attcgctaga
     3061 ttttagttag aattgcctga ttccacaccc ttcttagttt ttttcaatga gatgtatagt
     3121 ttatagtttt gcagaaaata aataaatttc atttaactcg cgaacatgtt gaagatatga
     3181 atattaatga gatgcgagta acattttaat ttgcagatgg ttgccatctt gattggcctc
     3241 atctttttgg gccaacaact cacgcaagtg ggcgtgatga atatcaacgg agccatcttc
     3301 ctcttcctga ccaacatgac ctttcaaaac gtctttgcca cgataaatgt aagtcttgtt
     3361 tagaatacat ttgcatatta ataatttact aactttctaa tgaatcgatt cgatttaggt
     3421 gttcacctca gagctgccag tttttatgag ggaggcccga agtcgacttt atcgctgtga
     3481 cacatacttt ctgggcaaaa cgattgccga attaccgctt tttctcacag tgccactggt
     3541 cttcacggcg attgcctatc cgatgatcgg actgcgggcc ggagtgctgc acttcttcaa
     3601 ctgcctggcg ctggtcactc tggtggccaa tgtgtcaacg tccttcggat atctaatatc
     3661 ctgcgccagc tcctcgacct cgatggcgct gtctgtgggt ccgccggtta tcataccatt
     3721 cctgctcttt ggcggcttct tcttgaactc gggctcggtg ccagtatacc tcaaatggtt
     3781 gtcgtacctc tcatggttcc gttacgccaa cgagggtctg ctgattaacc aatgggcgga
     3841 cgtggagccg ggcgaaatta gctgcacatc gtcgaacacc acgtgcccca gttcgggcaa
     3901 ggtcatcctg gagacgctta acttctccgc cgccgatctg ccgctggact acgtgggtct
     3961 ggccattctc atcgtgagct tccgggtgct cgcatatctg gctctaagac ttcgggcccg
     4021 acgcaaggag tagccgacat atatccgaaa taactgcttg tttttttttt ttaccattat
     4081 taccatcgtg tttactgttt attgccccct caaaaagcta atgtaattat atttgtgcca
     4141 ataaaaacaa gatatgacct atagaataca agtatttccc cttcgaacat ccccacaagt
     4201 agactttgga tttgtcttct aaccaaaaga cttacacacc tgcatacctt acatcaaaaa
     4261 ctcgtttatc gctacataaa acaccgggat atatttttta tatacatact tttcaaatcg
     4321 cgcgccctct tcataattca cctccaccac accacgtttc gtagttgctc tttcgctgtc
     4381 tcccacccgc tctccgcaac acattcacct tttgttcgac gaccttggag cgactgtcgt
     4441 tagttccgcg cgattcggtt cgctcaaatg gttccgagtg gttcatttcg tctcaataga
     4501 aattagtaat aaatatttgt atgtacaatt tatttgctcc aatatatttg tatatatttc
     4561 cctcacagct atatttattc taatttaata ttatgacttt ttaaggtaat tttttgtgac
     4621 ctgttcggag tgattagcgt tacaatttga actgaaagtg acatccagtg tttgttcctt
     4681 gtgtagatgc atctcaaaaa aatggtgggc ataatagtgt tgtttatata tatcaaaaat
     4741 aacaactata ataataagaa tacatttaat ttagaaaatg cttggatttc actggaacta
     4801 ggctagattt aaatgcatgc cctgcaggaa gcttatgatc ctgacgacgg agaccgccgt
     4861 cgtcgacaag cccctaggag gcctagatct gcggccgctt aattaaacgc gtgaattcac
     4921 cggtccgcgg ggcgcgccgc tagccatatg ggtaccggat ccggcttgtc gacgacggcg
     4981 gtctccgtcg tcaggatcat actagttcta gccagctttt gttcccttta gtgagggtta
     5041 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5101 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5161 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5641 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5881 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7201 cgaaaagtgc cac
//
