LOCUS       sequence-257089-        7780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the targeting of chloramphenicol gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7780)
  TITLE     BAC targeting vector (BTV)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7780)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CTV
FEATURES             Location/Qualifiers
     source          1..7780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..57
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     49..66
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     50..66
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     73..92
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        73..91
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(631..793,794..805)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             complement(join(870..1661,1662..1730))
                     /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"
     promoter        complement(1731..1834)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(1858..1869,1870..2057)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    2410..2531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2421..2440)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2475..2494
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2705..3499)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRTAFYRLLDEFF"
     promoter        complement(3566..3895)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3580..3715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3634..3653)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3971..3990
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4048..4066)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4049..4066)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4084..4100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4084..4100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4097..4119)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4108..4124
                     /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(4132..4138,4139..4156,4157..4162))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4177..4198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4315..4332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4486..5074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4566..4585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5245..6036,6037..6105))
                     /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"
     CDS             complement(6123..6917)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(7261..7689)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7371..7392)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7583..7602
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
       61 gccagtgaat tgtaatacga ctcactatag ggcgaattgg gccctctaga tgcatgctcg
      121 agcggccgcc agtgtgatgg atatattcat taagcattct gccgacatgg aagccatcac
      181 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
      241 atttgatatc tgcagaattc gcccttccca tctggcctgt gtttcagaca ccagggagtc
      301 tctgctcacg tttcctgcta tttgcagcct ctctatcaag actaatacac ctcttcccgc
      361 atcggctgcc tgtgagaggc ttttcagcac tgcaggattg cttttcagcc ccaaaagagc
      421 taggcttgac actaacaatt ttgagaatca gcttctactg aagttaaatc tgaggtttta
      481 caactttgag tagcgtgtac tggcattaga ttgtctgtct tatagtttga taattaaata
      541 caaacagttc taaagcagga taaaaccttg tatgcatttc atttaatgtt ttttgagatt
      601 aaaagcttaa acaagaatct ctagttttct ttcttgcttt tacttttact tccttaatac
      661 tcaagtacaa ttttaatgga gtactttttt acttttactc aagtaagatt ctagccagat
      721 acttttactt ttaattgagt aaaattttcc ctaagtactt gtactttcac ttgagtaaaa
      781 tttttgagta ctttttacac ctctgtcaag aactcctgga caaacctctg acctgtgtgg
      841 aacagagtgg atatgggttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      901 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      961 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1021 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1081 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1141 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1201 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1261 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1321 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1381 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1441 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1501 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1561 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1621 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1681 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1741 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1801 tgtatttaga aaaataaaca aataggggtt ccgcaatgaa ggtagggggt caagaaccag
     1861 aggtgtaaag tacttgagta attttacttg attactgtac ttaagtatta tttttgggga
     1921 tttttacttt acttgagtac aattaaaaat caatactttt acttttactt aattacattt
     1981 ttttagaaaa aaaagtactt tttactcctt acaattttat ttacagtcaa aaagtactta
     2041 ttttttggag atcacttcat tctattttcc cttgctatta ccaaaccaat tgaattgcgc
     2101 tgatgcccag tttaatttaa atgttattta ttctgcctat gaaaatcgtt ttcacattat
     2161 atgaaattgg tcagacatgt tcattggtcc tttggaagtg acgtcatgtc acatctatta
     2221 ccacaatgca cagcaccttg acctggaaat tagggaaatt ataacagtca atcagtggaa
     2281 gaaaatggag gaagtatgtg attcatcagc agctgcgagc agcacagtcc aaaatcagcc
     2341 acaggatcaa gagcacccgt ggccgtatct tcgaagggcg aattccagca cactggcggc
     2401 cgttactagt aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2461 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2521 ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     2581 cagccggcgt cccggaaaac gattccgaag cccgaccttt catagaaggc ggcggtggaa
     2641 tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     2701 ccgctcagaa gaactcgtca agaaggcgat agaaggcggt gcgctgcgaa tcgggagcgg
     2761 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     2821 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     2881 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     2941 tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg
     3001 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     3061 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     3121 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     3181 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     3241 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     3301 gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     3361 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     3421 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     3481 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatccgaaaa
     3541 tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     3601 actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa
     3661 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     3721 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     3781 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     3841 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacctagt
     3901 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     3961 ggtacattga gcaactgact gaaatgcctc aaaatgttct ctagtggatc cgagctcggt
     4021 accaagcttg atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
     4081 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4141 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4321 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4381 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4441 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4501 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4561 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4621 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4681 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4741 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4801 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4861 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4921 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4981 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5041 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5101 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5161 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5221 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5281 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5341 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5401 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5461 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5521 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5581 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5641 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5701 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5761 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5821 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5881 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5941 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6001 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6061 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6121 attcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     6181 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     6241 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     6301 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     6361 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     6421 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     6481 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     6541 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     6601 tgggatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     6661 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     6721 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     6781 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     6841 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     6901 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     6961 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     7021 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc
     7081 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg
     7141 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     7201 ggactggctt tctacgtgtt ccgcttcctt tagcagccct tgcgccctga attttgttaa
     7261 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     7321 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     7381 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     7441 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     7501 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7561 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7621 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7681 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7741 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
//
