LOCUS       sequence-252296-        7781 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 7781)
  TITLE     BAC targeting vector (BTV)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7781)
  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            "Unpublished"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..7781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(315..334)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        410..739
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      590..725
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     652..671
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             806..1600
                     /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"
     polyA_signal    1774..1895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1865..1884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(2249..2436,2437..2448)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        2472..2575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2576..2644,2645..3436)
                     /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"
     misc_feature    join(3501..3512,3513..3675)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4214..4233)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4215..4233)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4240..4257)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4240..4256)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4249..4271)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4397..4825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4484..4503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4694..4715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             5169..5963
                     /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"
     CDS             join(5981..6049,6050..6841)
                     /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"
     rep_origin      7012..7600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7501..7520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7754..7771
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
      361 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagactagg tgtgtcagtt
      421 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      481 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      541 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
      601 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
      661 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
      721 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
      781 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
      841 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
      901 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
      961 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     1021 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     1081 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     1141 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     1201 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     1261 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     1321 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     1381 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     1441 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1501 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1561 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     1621 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     1681 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1741 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     1801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1861 agttgtggtt tgtccaaact catcaatgta tcttactagt aacggccgcc agtgtgctgg
     1921 aattcgccct tgcgaagata cggccacggg tgctcttgat cctgtggctg attttggact
     1981 gtgctgctcg cagctgctga tgaatcacat acttcctcca ttttcttcca ctgattgact
     2041 gttataattt ccctaatttc caggtcaagg tgctgtgcat tgtggtaata gatgtgacat
     2101 gacgtcactt ccaaaggacc aatgaacatg tctgaccaat ttcatataat gtgaaaacga
     2161 ttttcatagg cagaataaat aacatttaaa ttaaactggg catcagcgca attcaattgg
     2221 tttggtaata gcaagggaaa atagaatgaa gtgatctcca aaaaataagt actttttgac
     2281 tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg taattaagta
     2341 aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc aaaaataata
     2401 cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggt tcttgacccc
     2461 ctaccttcat tgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2521 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2581 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2641 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2701 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2761 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2821 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2881 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2941 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3001 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3061 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3121 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3181 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3241 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3301 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3361 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3421 gattaagcat tggtaactgt cagaccaacc catatccact ctgttccaca caggtcagag
     3481 gtttgtccag gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa
     3541 agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct
     3601 tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa
     3661 aagtaaaagc aagaaagaaa actagagatt cttgtttaag cttttaatct caaaaaacat
     3721 taaatgaaat gcatacaagg ttttatcctg ctttagaact gtttgtattt aattatcaaa
     3781 ctataagaca gacaatctaa tgccagtaca cgctactcaa agttgtaaaa cctcagattt
     3841 aacttcagta gaagctgatt ctcaaaattg ttagtgtcaa gcctagctct tttggggctg
     3901 aaaagcaatc ctgcagtgct gaaaagcctc tcacaggcag ccgatgcggg aagaggtgta
     3961 ttagtcttga tagagaggct gcaaatagca ggaaacgtga gcagagactc cctggtgtct
     4021 gaaacacagg ccagatggga agggcgaatt ctgcagatat caaatattat acgcaaggcg
     4081 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     4141 tcggcagaat gcttaatgaa tatatccatc acactggcgg ccgctcgagc atgcatctag
     4201 agggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     4261 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4321 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4381 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4441 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4501 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4561 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4621 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4681 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4741 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4801 atttaacaaa aatttaacgc gaattttaac aaaattcagg gcgcaagggc tgctaaagga
     4861 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     4921 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
     4981 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
     5041 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
     5101 tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat
     5161 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5221 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5281 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5341 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5401 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5461 cggggcagga tctcctgtca tcccaccttg ctcctgccga gaaagtatcc atcatggctg
     5521 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5581 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5641 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5701 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5761 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5821 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5881 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5941 gccttcttga cgagttcttc tgaattgaaa aaggaagagt atgagtattc aacatttccg
     6001 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6061 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6121 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6181 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6241 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6301 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6361 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6421 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6481 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6541 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6601 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6661 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6721 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6781 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6841 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6901 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6961 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7021 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7081 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7141 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7201 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7261 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7321 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7381 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7441 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7501 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7561 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7621 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7681 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7741 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
