LOCUS       sequence-139149-        7302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pTARBAC2 with kanaR selection marker and 
            lens-specific transgenesis marker Cerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  AUTHORS   Fuentes F, Reynolds E, Lewellis SW, Venkiteswaran G, Knaut H
  TITLE     A Plasmid Set for Efficient Bacterial Artificial Chromosome (BAC) 
            Transgenesis in Zebrafish.
  JOURNAL   G3 (Bethesda). 2016 Jan 27. pii: g3.115.026344. doi: 
            10.1534/g3.115.026344.
  PUBMED    26818072
REFERENCE   2  (bases 1 to 7302)
  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: "G3 
            (Bethesda). 2016 Jan 27. pii: g3.115.026344. doi: 
            10.1534/g3.115.026344."
COMMENT     Sequence Label: pBS-TarHom-Tol2-kanaR-cryaa-CFP
FEATURES             Location/Qualifiers
     source          1..7302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..101
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     93..110
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     94..110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        120..138
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(759..921,922..933)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1367..1386)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1378..2193
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGELTGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1451..1470)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    join(2583..2594,2595..2782)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     regulatory      3820..3829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3826..3828,3829..3831,3832..4545)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3871..3892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4132..4151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4479..4500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4670..4791
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4707..4726)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4761..4780
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5114..5134)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5114..5132)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5153..5169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5153..5169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5166..5188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5177..5193
                     /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(5201..5207,5208..5225,5226..5231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5246..5267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5384..5401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5555..6143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5635..5654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6314..7105,7106..7174))
                     /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     6937..6956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7175..7279)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
       61 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
      121 aatacgactc actatagggc gaattggagc tcggcggccg ccaggcctac ccactagtca
      181 attcgggagg atcgaaacgg cagatcgcaa aaaacagtac atacagaagg agacatgaac
      241 atgaacatca aaaaaattgt aaaacaagcc acagttctga cttttacgac tgcacttctg
      301 gcaggaggag cgactcaagc cttcgcgaaa gaaaataacc aaaaagcata caaagaaacg
      361 tacgcgtctc tcatattaca cgccgcggtg gcggccgcat ctggccttgt ttcagacacc
      421 agggagtctc tgctcacgtt tcctgctatt tgcagcctct ctatcaagac taatacacct
      481 cttcccgcat cggctgcctg tgagaggctt ttcagcactg cgggattgct tttcagcccc
      541 aaaagagcta ggcttgacac taacaatttt gagaatcagc ttctactgaa gttaaatctg
      601 aggttttaca actttgagta gcgtgtactg gcgttagatt gtctgtctta tagtttgata
      661 attaaataca aacagttcta aagcaggata aaaccttgta tgcatttcat ttaatgtttt
      721 ttgagattaa aagcttaaac aagaatctct agttttcttt cttgctttta cttttacttc
      781 cttaatactc aagtacaatt ttaatggagt acttttttac ttttactcaa gtaagattct
      841 agccagatac ttttactttt aattgagtaa aattttccct aagtacttgt actttcactt
      901 gagtaaaatt tttgagtact ttttacacct ctgtcaagaa ctcctggaca aacctctgac
      961 ctgtgtggaa cagagtggat atgggtgtct gaacagatat tcacgtcttt tgcagatcag
     1021 agggcatttc tggtgttgtg caatgacctg ggtccaacag gtaggcgtac tcgtatccaa
     1081 gagcattgtt ggacaggagg tagtctccat tcctataaag aggaaggaat ggcaccatgt
     1141 agtagccgtc attgtggcca attggtgcat tggtacgtgg gtagatggac cggggaggct
     1201 gatgagttct tagccagcgg atctgattac cctgttatcc ctagatttaa atgatatcgg
     1261 atcctagtaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1321 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     1381 agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct
     1441 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1501 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1561 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1621 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1681 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1741 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1801 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1861 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1921 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1981 gataacctta tttttgacga gggggaatta acaggttgta ttgatgttgg acgagtcgga
     2041 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2101 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2161 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2221 tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaataaat cgaactcttg
     2281 ctgagttgaa ggatcggatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc
     2341 aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc
     2401 tcactttctg gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt
     2461 cacgaggcag acctcagcgg cggccggatc cgatatcgtt tggcgcgcca aactcgctac
     2521 cttaggaccg ttatagttaa gatccgatct gctgggcttg ctgaaggtag ggggtcaaga
     2581 accagaggtg taaagtactt gagtaatttt acttgattac tgtacttaag tattattttt
     2641 ggggattttt actttacttg agtacaatta aaaatcaata cttttacttt tacttaatta
     2701 cattttttta gaaaaaaaag tactttttac tccttacaat tttatttaca gtcaaaaagt
     2761 acttattttt tggagatcac ttcattctat tttcccttgc tattaccaaa ccaattgaat
     2821 tgcgctgatg cccagtttaa tttaaatgtt atttattctg cctatgaaaa tcgttttcac
     2881 attatatgaa attggtcaga catgttcatt ggtcctttgg aagtgacgtc atgtcacatc
     2941 tattaccaca atgcacagca ccttgacctg gaaattaggg aaattataac agtcaatcag
     3001 tggaagaaaa tggaggaagt atgtgattca tcagcagctg cgagcagcac agtccaaaat
     3061 cagccacagg atcaagagca cccgtggccg tatcttcgcc ccgggagctt cgaattctaa
     3121 tatggcaata aacgatcttc agagaaactt gcattggtaa atagtttcag cctatgcata
     3181 tgtagttatt caaactgtaa tgactatctt acagtattat atgtttataa tagtttgatc
     3241 caataaatcc agatttatat gcttttcaac ataaaattgt gaaatgcaca gaatgtacat
     3301 atacaataca tttcctacaa taatgctctc ttttctatta atagtgtgca ttcagtgcag
     3361 gatgttatgt atttattttc tgtagtgcaa actttttcaa agaattttcc agtctaagct
     3421 tttcagtcaa gaaaaaaaaa agcgattgat tatctgtgat atttaaaaaa attagctctt
     3481 aaatctggtg aaccaagtgg ccacaacaca ttaaatactg acgttctaca gccatgtagc
     3541 ttcatctggt ctggtttgct ttggcaggca cttattttcc ggtgatagtt ggccacaaga
     3601 gtattgtctg agccattcag tgctagactg tcattctcag gtcagagtcc accccgctga
     3661 ttctgctgac aacactgttc ccaccatgag ataatgccat tccagagaga tccatttgta
     3721 agcccctctt tctgcagcac aggtatataa ccaggggtct gcctccacta aggccggcac
     3781 acatcatttg gggatctttg tactgtggat ccaccggtcg ccaccatggt gagcaagggc
     3841 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3901 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3961 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     4021 acctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     4081 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     4141 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     4201 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4261 gccatcagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     4321 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4381 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4441 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4501 accgccgccg ggatcactct cggcatggac gagctgtaca agtagcggcc ggccgcgact
     4561 ctagatcata atcagccata ccatatttgt agaggtttta cttgctttaa aaaacctccc
     4621 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4681 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4741 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aatcgagggg
     4801 gggcccggta cccaattcgc cctatagtga gtctcgagcc aagtgcctca attcgatcaa
     4861 tcaacgatta aaaatattga gtctgcaaaa ggacttgatg tgtgggacag ctggccgctg
     4921 caaaacgctg acggaacagt agctgaatac aacggctatc acgttgtgtt tgctcttgcg
     4981 ggaagcccga aagacgctga tgacacatca atctacatgt tttatcaaaa ggtcggcgac
     5041 aactcaatcg acagctggaa aaacgcgggc cgtgtcttta ctcgaggggg ggcccggtac
     5101 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     5161 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5221 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5281 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5341 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5401 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5461 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5521 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5581 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5761 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5881 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5941 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6001 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6061 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6121 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6181 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6241 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6301 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6361 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6421 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6481 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6541 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6601 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6661 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6721 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6781 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6841 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6901 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6961 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7021 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7081 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7141 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7201 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7261 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac
//
