LOCUS       sequence-140684-        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pTARBAC2 with kanaR selection marker and 
            lens-specific transgenesis marker sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  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 7301)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..7301
                     /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(2582..2593,2594..2781)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     regulatory      3819..3828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3825..3827,3828..3830,3831..4544)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3870..3891)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4478..4499
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4669..4790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4706..4725)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4760..4779
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5113..5133)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5113..5131)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5152..5168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5152..5168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5165..5187)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5176..5192
                     /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(5200..5206,5207..5224,5225..5230))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5245..5266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5383..5400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5554..6142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5634..5653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6313..7104,7105..7173))
                     /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     6936..6955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7174..7278)
                     /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 gagccttgtt 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 ttatattaag atccgatctg ctgggcttgc tgaaggtagg gggtcaagaa
     2581 ccagaggtgt aaagtacttg agtaatttta cttgattact gtacttaagt attatttttg
     2641 gggattttta ctttacttga gtacaattaa aaatcaatac ttttactttt acttaattac
     2701 atttttttag aaaaaaaagt actttttact ccttacaatt ttatttacag tcaaaaagta
     2761 cttatttttt ggagatcact tcattctatt ttcccttgct attaccaaac caattgaatt
     2821 gcgctgatgc ccagtttaat ttaaatgtta tttattctgc ctatgaaaat cgttttcaca
     2881 ttatatgaaa ttggtcagac atgttcattg gtcctttgga agtgacgtca tgtcacatct
     2941 attaccacaa tgcacagcac cttgacctgg aaattaggga aattatatca gtcaatcagt
     3001 ggaagaaaat ggaggaagta tgtgattcat cagcagctgc gagcagcaca gtccaaaatc
     3061 agccacagga tcaagagcac ccgtggccgt atcttcgccc cgggagcttc gaattctaat
     3121 atggcaataa acgatcttca gagaaacttg cattggtaaa tagtttcagc ctatgcatat
     3181 gtagttattc aaactgtaat gactatctta cagtattata tgtttataat agtttgatcc
     3241 aataaatcca gatttatatg cttttcaaca taaaattgtg aaatgcacag aatgtacata
     3301 tacaatacat ttcctacaat aatgctctct tttctattaa tagtgtgcat tcagtgcagg
     3361 atgttatgta tttattttct gtagtgcaaa ctttttcaaa gaattttcca gtctaagctt
     3421 ttcagtcaag aaaaaaaaaa gcgattgatt atctgtgata tttaaaaaaa ttagctctta
     3481 aatctggtga accaagtggc cacaacacat taaatactga cgttctacag ccatgtagct
     3541 tcatctggtc tggtttgttt tggcaggcac ttattttccg gtgatagttg gccacaagag
     3601 tattgtctga gccattcagt gctagactgt cattctcagg tcagagtcca ccccgctgat
     3661 tctgctgaca acactgttcc caccatgaga taatgccatt ccagagagat ccatttgtaa
     3721 gcccctcttt ctgcagcaca ggtatataac caggggtctg cctccactaa ggccggcaca
     3781 catcatttgg ggatctttgt actgtggatc caccggtcgc caccatggtg agcaagggcg
     3841 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3901 acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     3961 agttcatctg caccaccggc aagctgcccg tgccttggcc caccctcgtg accaccctga
     4021 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcgccac gacttcttca
     4081 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag gacgacggca
     4141 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4201 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4261 tcaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     4321 tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4381 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4441 ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4501 ccgccgccgg gatcactcac ggcatggacg agctgtacaa gtagcggccg gccgcgactc
     4561 tagatcataa tcagccatac catatttgta gaggttttac ttgctttaaa aaacctccca
     4621 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4681 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4741 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta atcgaggggg
     4801 ggcccggtac ccaattcgcc ctatagtgag tctcgagcca agtgcctcaa ttcgatcaat
     4861 caacgattaa aaatattgag tctgcaaaag gacttgatgt gtgggacagc tggccgctgc
     4921 aaaacgctga cggaacagta gctgaataca acggctatca cgttgtgttt gctcttgcgg
     4981 gaagcccgaa agacgctgat gacacatcaa tctacatgtt ttatcaaaag gtcggcgaca
     5041 actcaatcga cagctggaaa aacgcgggcc gtgtctttac tcgagggggg gcccggtacc
     5101 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     5161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5221 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5641 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6361 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6721 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7261 acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
