LOCUS       sequence-393499-        7838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pTARBAC2 with GalK selection marker and 
            lens-specific transgenesis marker Cerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7838)
  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 7838)
  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-FRT-GalK-cryaa-CFP
FEATURES             Location/Qualifiers
     source          1..7838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /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     complement(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    join(2807..2969,2970..2981)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    3283..3316
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     3325..3341
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3325..3341
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4523..4570)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    4574..4607
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    join(4662..4673,4674..4861)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     regulatory      5899..5908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5905..5907,5908..5910,5911..6624)
                     /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(5950..5971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6211..6230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6558..6579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6749..6870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6786..6805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6840..6859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(7192..7210)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      7378..7806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7465..7484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7675..7696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ssmcwttymg gskrmryrwg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctykrwaw tamsmctcac tatagggcga attggagctc ggcggccgcc aggcctaccc
     2221 actagtcaat tcgggaggat cgaaacggca gatcgcaaaa aacagtacat acagaaggag
     2281 acatgaacat gaacatcaaa aaaattgtaa aacaagccac agttctgact tttacgactg
     2341 cacttctggc aggaggagcg actcaagcct tcgcgaaaga aaataaccaa aaagcataca
     2401 aagaaacgta cgcgtctctc atattacacg ccgcggtggc ggccgcatct ggccttgttt
     2461 cagacaccag ggagtctctg ctcacgtttc ctgctatttg cagcctctct atcaagacta
     2521 atacacctct tcccgcatcg gctgcctgtg agaggctttt cagcactgcg ggattgcttt
     2581 tcagccccaa aagagctagg cttgacacta acaattttga gaatcagctt ctactgaagt
     2641 taaatctgag gttttacaac tttgagtagc gtgtactggc gttagattgt ctgtcttata
     2701 gtttgataat taaatacaaa cagttctaaa gcaggataaa accttgtatg catttcattt
     2761 aatgtttttt gagattaaaa gcttaaacaa gaatctctag ttttctttct tgcttttact
     2821 tttacttcct taatactcaa gtacaatttt aatggagtac ttttttactt ttactcaagt
     2881 aagattctag ccagatactt ttacttttaa ttgagtaaaa ttttccctaa gtacttgtac
     2941 tttcacttga gtaaaatttt tgagtacttt ttacacctct gtcaagaact cctggacaaa
     3001 cctctgacct gtgtggaaca gagtggatat gggtgtctga acagatattc acgtcttttg
     3061 cagatcagag ggcatttctg gtgttgtgca atgacctggg tccaacaggt aggcgtactc
     3121 gtatccaaga gcattgttgg acaggaggta gtctccattc ctataaagag gaaggaatgg
     3181 caccatgtag tagccgtcat tgtggccaat tggtgcattg gtacgtgggt agatggaccg
     3241 gggaggctga tgagttctta gccagcggat cttggttagt tagaagttcc tattctctag
     3301 aaagtatagg aacttcgcgg ccgctctaga actagtggat cctcagcact gtcctgctcc
     3361 ttgtgatggt ttacaaacgt aaaaagtctc tttaatacct gtttttgctt catattgttc
     3421 agcgacagct tgctgtacgg caggcaccag ctcttccggg atcagcgcga cgatacagcc
     3481 gccaaatccg ccgccggtca tgcgtacgcc acctttgtcg ccaatcacag ctttgacgac
     3541 ttctaccaga gtgtcaattt gcggcacggt gatttcgaaa tcatcgcgca tagaggcatg
     3601 agactccgcc atcaactcgc ccatacgttt caggtcgcct tgctccagcg cgctggcagc
     3661 ttcaacggtg cgggcgtttt cagtcagtat atgacgcacg cgttttgcca cgatcgggtc
     3721 cagttcatgc gcaacagcgt tgaactcttc aatggtgaca tcacgcaggg ctggctgctg
     3781 gaagaaacgc gcaccggttt cgcactgttc acgacgggtg ttgtattcgc tgccaaccag
     3841 ggtacgtttg aagttactgt tgatgatgac gacagccaca cctttgggca tggaaactgc
     3901 tttggtcccc agtgagcggc aatcgatcag caaggcatga tctttcttgc cgagcgcgga
     3961 aattagctga tccatgatcc cgcagttaca gcctacaaac tggttttctg cttcctgacc
     4021 gttaagcgcg atttgtgcgc cgtccagcgg gagatgataa agctgctgca atacggttcc
     4081 gaccgcgact tccagtgaag cggaagaact taacccggca ccctgcggca cattgccgct
     4141 gatcaccatg tccacgccgc cgaagctgtt gttacgcagt tgcagatgtt tcaccacgcc
     4201 acgaacgtag ttagcccatt gatagttttc atgtgcgaca atgggcgcat cgagggaaaa
     4261 ctcgtcgagc tgattttcat aatcggctgc catcacgcga actttacggt catcgcgtgg
     4321 tgcacaactg atcacggttt gataatcaat cgcgcagggc agaacgaaac cgtcgttgta
     4381 gtcggtgtgt tcaccaatca aattcacgcg gccaggcgcc tgaatggtgt gagtggcagg
     4441 gtagccaaat gcgttggcaa acagagattg tgttttttct ttcagactca tgatctgcct
     4501 cctgggttta gtccctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt
     4561 aattgtcaac agggaagttc ctattctcta gaaagtatag gaacttcgta ccgaactgag
     4621 atccgatctg ctgggcttgc tgaaggtagg gggtcaagaa ccagaggtgt aaagtacttg
     4681 agtaatttta cttgattact gtacttaagt attatttttg gggattttta ctttacttga
     4741 gtacaattaa aaatcaatac ttttactttt acttaattac atttttttag aaaaaaaagt
     4801 actttttact ccttacaatt ttatttacag tcaaaaagta cttatttttt ggagatcact
     4861 tcattctatt ttcccttgct attaccaaac caattgaatt gcgctgatgc ccagtttaat
     4921 ttaaatgtta tttattctgc ctatgaaaat cgttttcaca ttatatgaaa ttggtcagac
     4981 atgttcattg gtcctttgga agtgacgtca tgtcacatct attaccacaa tgcacagcac
     5041 cttgacctgg aaattaggga aattatatca gtcaatcagt ggaagaaaat ggaggaagta
     5101 tgtgattcat cagcagctgc gagcagcaca gtccaaaatc agccacagga tcaagagcac
     5161 ccgtggccgt atcttcgccc cgggagcttc gaattctaat atggcaataa acgatcttca
     5221 gagaaacttg cattggtaaa tagtttcagc ctatgcatat gtagttattc aaactgtaat
     5281 gactatctta cagtattata tgtttataat agtttgatcc aataaatcca gatttatatg
     5341 cttttcaaca taaaattgtg aaatgcacag aatgtacata tacaatacat ttcctacaat
     5401 aatgctctct tttctattaa tagtgtgcat tcagtgcagg atgttatgta tttattttct
     5461 gtagtgcaaa ctttttcaaa gaattttcca gtctaagctt ttcagtcaag aaaaaaaaaa
     5521 gcgattgatt atctgtgata tttaaaaaaa ttagctctta aatctggtga accaagtggc
     5581 cacaacacat taaatactga cgttctacag ccatgtagct tcatctggtc tggtttgctt
     5641 tggcaggcac ttattttccg gtgatagttg gccacaagag tattgtctga gccattcagt
     5701 gctagactgt cattctcagg tcagagtcca ccccgctgat tctgctgaca acactgttcc
     5761 caccatgaga taatgccatt ccagagagat ccatttgtaa gcccctcttt ctgcagcaca
     5821 ggtatataac caggggtctg cctccactaa ggccggcaca catcatttgg ggatctttgt
     5881 actgtggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     5941 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     6001 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     6061 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
     6121 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     6181 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     6241 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     6301 gaggacggca acatcctggg gcacaagctg gagtacaacg ccatcagcga caacgtctat
     6361 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     6421 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     6481 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     6541 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     6601 ggcatggacg agctgtacaa gtagcggccg gccgcgactc tagatcataa tcagccatac
     6661 catatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     6721 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     6781 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6841 tggtttgtcc aaactcatca atgtatctta atcgaggggg ggcccggtac ccaattcgcc
     6901 ctatagtgag tctcgagcca agtgcctcaa ttcgatcaat caacgattaa aaatattgag
     6961 tctgcaaaag gacttgatgt gtgggacagc tggccgctgc aaaacgctga cggaacagta
     7021 gctgaataca acggctatca cgttgtgttt gctcttgcgg gaagcccgaa agacgctgat
     7081 gacacatcaa tctacatgtt ttatcaaaag gtcggcgaca actcaatcga cagctggaaa
     7141 aacgcgggcc gtgtctttac tcgagggggg gcccggtacc agcttttgtt ccctttagtg
     7201 agggttaatt tcgagcttgg cgtaatcatg gwcawmgyyg tkwcnnntgg gaaaaysytg
     7261 kcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     7321 aagaggcccg caccgatcgc ccttcccaac agtygckcms cckraawgks smayskgack
     7381 ygccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     7441 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     7501 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     7561 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     7621 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     7681 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     7741 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     7801 cgaattttaa caaaatatta acgcttacaa ktymsrtk
//
