LOCUS       sequence-139151-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pTARBAC2 with kanaR selection marker and 
            lens-specific transgenesis marker dsRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  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 7259)
  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-dsRed
FEATURES             Location/Qualifiers
     source          1..7259
                     /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             3826..4503
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MAPSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPAEIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(3956..3974)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(4006..4026)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4224..4243
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4418..4441
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4628..4749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4665..4684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4719..4738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5071..5091)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5071..5089)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5110..5126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5110..5126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5123..5145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5134..5150
                     /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(5158..5164,5165..5182,5183..5188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5203..5224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5341..5358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5512..6100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5592..5611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6271..7062,7063..7131))
                     /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     6894..6913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7132..7236)
                     /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 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 ctggtttgtt 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 ccaccatggc cccctccgag
     3841 gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
     3901 gaattcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3961 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     4021 cagtacggct ccaaggtgta cgtgaagcac cccgccgaaa tccccgacta caagaagctg
     4081 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     4141 accgtgaccc aggactcctc cctgcaggac ggctccttca tctacaaggt gaagttcatc
     4201 ggcgtgaact tcccctccga cggccccgta atgcagaaga agactatggg ctgggaggcc
     4261 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca caaggccctg
     4321 aagctgaagg acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag
     4381 cccgtgcagc tgcccggcta ctactacgtg gactccaagc tggacatcac ctcccacaac
     4441 gaggactaca ccatcgtgga gcagtacgag cgcgccgagg gccgccacca cctgttcctg
     4501 tagcggccgg ccgcgactct agatcataat cagccatacc atatttgtag aggttttact
     4561 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4621 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4741 tgtatcttaa tcgagggggg gcccggtacc caattcgccc tatagtgagt ctcgagccaa
     4801 gtgcctcaat tcgatcaatc aacgattaaa aatattgagt ctgcaaaagg acttgatgtg
     4861 tgggacagct ggccgctgca aaacgctgac ggaacagtag ctgaatacaa cggctatcac
     4921 gttgtgtttg ctcttgcggg aagcccgaaa gacgctgatg acacatcaat ctacatgttt
     4981 tatcaaaagg tcggcgacaa ctcaatcgac agctggaaaa acgcgggccg tgtctttact
     5041 cgaggggggg cccggtacca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     5101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5941 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
