LOCUS       sequence-139599-        8418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pTARBAC2 with tetR selection marker and 
            lens-specific transgenesis marker Citrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8418)
  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 8418)
  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-TetR-cryaa-YFP
FEATURES             Location/Qualifiers
     source          1..8418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(1267..1429,1430..1441)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     promoter        join(1742..1747,1748..1764,1765..1770)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1818..3008
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(2170..2189)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    join(3696..3707,3708..3895)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     regulatory      4933..4942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4943..4945,4946..4948,4949..5659)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4988..5009)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5249..5268)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5596..5617
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5786..5907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5823..5842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5877..5896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6230..6250)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6230..6248)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6269..6285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6269..6285)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6282..6304)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6293..6309
                     /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(6317..6323,6324..6341,6342..6347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6362..6383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6500..6517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6671..7259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6751..6770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7430..8221,8222..8290))
                     /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     8053..8072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8291..8395)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcgg
      661 cggccgccag gcctacccac tagtcaattc gggaggatcg aaacggcaga tcgcaaaaaa
      721 cagtacatac agaaggagac atgaacatga acatcaaaaa aattgtaaaa caagccacag
      781 ttctgacttt tacgactgca cttctggcag gaggagcgac tcaagccttc gcgaaagaaa
      841 ataaccaaaa agcatacaaa gaaacgtacg gcgtctctca tattacacgc cgcggtggcg
      901 gccgcatctg gcctgtgttt cagacaccag ggagtctctg ctcacgtttc ctgctatttg
      961 cagcctctct atcaagacta atacacctct tcccgcatcg gctgcctgtg agaggctttt
     1021 cagcactgca ggattgcttt tcagccccaa aagagctagg cttgacacta acaattttga
     1081 gaatcagctt ctactgaagt taaatctgag gttttacaac tttgagtagc gtgtactggc
     1141 attagattgt ctgtcttata gtttgataat taaatacaaa cagttctaaa gcaggataaa
     1201 accttgtatg catttcattt aatgtttttt gagattaaaa gcttaaacaa gaatctctag
     1261 ttttctttct tgcttttact tttacttcct taatactcaa gtacaatttt aatggagtac
     1321 ttttttactt ttactcaagt aagattctag ccagatactt ttacttttaa ttgagtaaaa
     1381 ttttccctaa gtacttgtac tttcacttga gtaaaatttt tgagtacttt ttacacctct
     1441 gtcaagaact cctggacaaa cctctgacct gtgtggaaca gagtggatat gggtgtctga
     1501 acagatattc acgtcttttg cagatcagag ggcatttctg gtgttgtgca atgacctggg
     1561 tccaacaggt aggcgtactc gtatccaaga gccttgttgg acaggaggta gtctccattc
     1621 ctataaagag gaaggaatgg caccatgtag tagccgtcat tgtggccaat tggtgcattg
     1681 gtacgtgggt agatggaccg gggaggctga tgagttctta gccagcggat ctttctcatg
     1741 tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta aattgctaac
     1801 gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca
     1861 ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata
     1921 tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga
     1981 tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag
     2041 tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg
     2101 tcctgtggat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     2161 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     2221 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     2281 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     2341 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaccg atgcccttga
     2401 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     2461 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     2521 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     2581 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     2641 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     2701 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     2761 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     2821 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg
     2881 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
     2941 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
     3001 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
     3061 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     3121 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
     3181 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     3241 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     3301 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     3361 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
     3421 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
     3481 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
     3541 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
     3601 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccagatccga
     3661 tctgctgggc ttgctgaagg tagggggtca agaaccagag gtgtaaagta cttgagtaat
     3721 tttacttgat tactgtactt aagtattatt tttggggatt tttactttac ttgagtacaa
     3781 ttaaaaatca atacttttac ttttacttaa ttacattttt ttagaaaaaa aagtactttt
     3841 tactccttac aattttattt acagtcaaaa agtacttatt ttttggagat cacttcattc
     3901 tattttccct tgctattacc aaaccaattg aattgcgctg atgcccagtt taatttaaat
     3961 gttatttatt ctgcctatga aaatcgtttt cacattatat gaaattggtc agacatgttc
     4021 attggtcctt tggaagtgac gtcatgtcac atctattacc acaatgcaca gcaccttgac
     4081 ctggaaatta gggaaattat aacagtcaat cagtggaaga aaatggagga agtatgtgat
     4141 tcatcagcag ctgcgagcag cacagtccaa aatcagccac aggatcaaga gcacccgtgg
     4201 ccgtatcttc gccccgggag cttcgaattc taatatggca ataaacgatc ttcagagaaa
     4261 cttgcattgg taaatagttt cagcctatgc atatgtagtt attcaaactg taatgactat
     4321 cttacagtat tatatgttta taatagtttg atccaataaa tccagattta tatgcttttc
     4381 aacataaaat tgtgaaatgc acagaatgta catatacaat acatttccta caataatgct
     4441 ctcttttcta ttaatagtgt gcattcagtg caggatgtta tgtatttatt ttctgtagtg
     4501 caaacttttt caaagaattt tccagtctaa gcttttcagt caagaaaaaa aaaagcgatt
     4561 gattatctgt gatatttaaa aaaattagct cttaaatctg gtgaaccaag tggccacaac
     4621 acattaaata ctgacgttct acagccatgt agcttcatct ggtctggttt gctttggcag
     4681 gcacttattt tccggtgata gttggccaca agagtattgt ctgagccatt cagtgctaga
     4741 ctgtcattct caggtcagag tccaccccgc tgattctgct gacaacactg ttcccaccat
     4801 gagataatgc cattccagag agatccattt gtaagcccct ctttctgcag cacaggtata
     4861 taaccagggg tctgcctcca ctaaggccgg cacacatcat ttggggatct ttgtactgtg
     4921 gatccaccgg tcgccaccat ggatggtgag caagggcgag gagctgttca ccggggtggt
     4981 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5041 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5101 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc
     5161 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5221 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5281 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5341 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5401 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     5461 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     5521 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     5581 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     5641 catggacgag ctgtacaagt aaggccggcc gcgactctag atcataatca gccataccat
     5701 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5761 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5821 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5881 tttgtccaaa ctcatcaatg tatcttaatc gagggggggc ccggtaccca attcgcccta
     5941 tagtgagtct cgagccaagt gcctcaattc gatcaatcaa cgattaaaaa tattgagtct
     6001 gcaaaaggac ttgatgtgtg ggacagctgg ccgctgcaaa acgctgacgg aacagtagct
     6061 gaatacaacg gctatcacgt tgtgtttgct cttgcgggaa gcccgaaaga cgctgatgac
     6121 acatcaatct acatgtttta tcaaaaggtc ggcgacaact caatcgacag ctggaaaaac
     6181 gcgggccgtg tctttactcg agggggggcc cggtacccag cttttgttcc ctttagtgag
     6241 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6301 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6361 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6421 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6481 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6541 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6601 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6661 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6721 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6781 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6841 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6901 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6961 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7021 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7081 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7141 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7201 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7261 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7321 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7381 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7441 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7501 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7561 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7621 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7681 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7741 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7801 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7861 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7921 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7981 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8041 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8101 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8161 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8221 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8281 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8341 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8401 ttccccgaaa agtgccac
//
