LOCUS       sequence-134701-        7841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pTARBAC2 with GalK selection marker and 
            lens-specific transgenesis marker Citrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7841)
  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 7841)
  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-YFP
FEATURES             Location/Qualifiers
     source          1..7841
                     /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"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(2813..2975,2976..2987)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    3289..3322
                     /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     3331..3347
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3331..3347
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4529..4576)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    4580..4613
                     /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(4668..4679,4680..4867)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     regulatory      5905..5914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5915..5917,5918..5920,5921..6631)
                     /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(5960..5981)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6221..6240)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6568..6589
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6758..6879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6795..6814)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6849..6868
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(7198..7216)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7223..7240)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7223..7239)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7232..7254)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7381..7836
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7468..7487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7678..7699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcacttttcg gggaaatgtg 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 tgtccttcta 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 agctcgaaat taaccctcac taaagggaac aaaagctgga gctcggcggc cgccaggcct
     2221 acccactagt caattcggga ggatcgaaac ggcagatcgc aaaaaacagt acatacagaa
     2281 ggagacatga acatgaacat caaaaaaatt gtaaaacaag ccacagttct gacttttacg
     2341 actgcacttc tggcaggagg agcgactcaa gccttcgcga aagaaaataa ccaaaaagca
     2401 tacaaagaaa cgtacggcgt ctctcatatt acacgccgcg gtggcggccg catctggcct
     2461 gtgtttcaga caccagggag tctctgctca cgtttcctgc tatttgcagc ctctctatca
     2521 agactaatac acctcttccc gcatcggctg cctgtgagag gcttttcagc actgcaggat
     2581 tgcttttcag ccccaaaaga gctaggcttg acactaacaa ttttgagaat cagcttctac
     2641 tgaagttaaa tctgaggttt tacaactttg agtagcgtgt actggcatta gattgtctgt
     2701 cttatagttt gataattaaa tacaaacagt tctaaagcag gataaaacct tgtatgcatt
     2761 tcatttaatg ttttttgaga ttaaaagctt aaacaagaat ctctagtttt ctttcttgct
     2821 tttactttta cttccttaat actcaagtac aattttaatg gagtactttt ttacttttac
     2881 tcaagtaaga ttctagccag atacttttac ttttaattga gtaaaatttt ccctaagtac
     2941 ttgtactttc acttgagtaa aatttttgag tactttttac acctctgtca agaactcctg
     3001 gacaaacctc tgacctgtgt ggaacagagt ggatatgggt gtctgaacag atattcacgt
     3061 cttttgcaga tcagagggca tttctggtgt tgtgcaatga cctgggtcca acaggtaggc
     3121 gtactcgtat ccaagagcct tgttggacag gaggtagtct ccattcctat aaagaggaag
     3181 gaatggcacc atgtagtagc cgtcattgtg gccaattggt gcattggtac gtgggtagat
     3241 ggaccgggga ggctgatgag ttcttagcca gcggatcttg gttagttaga agttcctatt
     3301 ctctagaaag tataggaact tcgcggccgc tctagaacta gtggatcctc agcactgtcc
     3361 tgctccttgt gatggtttac aaacgtaaaa agtctcttta atacctgttt ttgcttcata
     3421 ttgttcagcg acagcttgct gtacggcagg caccagctct tccgggatca gcgcgacgat
     3481 acagccgcca aatccgccgc cggtcatgcg tacgccacct ttgtcgccaa tcacagcttt
     3541 gacgatttct accagagtgt caatttgcgg cacggtgatt tcgaaatcat cgcgcataga
     3601 ggcatgagac tccgccatca actcgcccat acgtttcagg tcgccttgct ccagcgcgct
     3661 ggcagcttca acggtgcggg cgttttcagt cagtatatga cgcacgcgtt ttgccacgat
     3721 cgggtccagt tcatgcgcaa cagcgttgaa ctcttcaatg gtgacatcac gcagggctgg
     3781 ctgctggaag aaacgcgcac cggtttcgca ctgttcacga cgggtgttgt attcgctgcc
     3841 aaccagggta cgtttgaagt tactgttgat gatgacgaca gccacacctt tgggcatgga
     3901 aactgctttg gtccccagtg agcggcaatc gatcagcaag gcatgatctt tcttgccgag
     3961 cgcggaaatt agctgatcca tgatcccgca gttacagcct acaaactggt tttctgcttc
     4021 ctgaccgtta agcgcgattt gtgcgccgtc cagcggcaga tgataaagct gctgcaatac
     4081 ggttccgacc gcgacttcca gtgaagcgga agaacttaac ccggcaccct gcggcacatt
     4141 gccgctgatc accatgtcca cgccgccgaa gctgttgtta cgcagttgca gatgtttcac
     4201 cacgccacga acgtagttag cccattgata gttttcatgt gcgacaatgg gcgcatcgag
     4261 ggaaaactcg tcgagctgat tttcataatc ggctgccatc acgcgaactt tacggtcatc
     4321 gcgtggtgca caactgatca cggtttgata atcaatcgcg cagggcagaa cgaaaccgtc
     4381 gttgtagtcg gtgtgttcac caatcaaatt cacgcggcca ggcgcctgaa tggtgtgagt
     4441 ggcagggtag ccaaatgcgt tggcaaacag agattgtgtt ttttctttca gactcatgat
     4501 ctgcctcctg ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg
     4561 atgattaatt gtcaacaggg aagttcctat tctctagaaa gtataggaac ttcgtaccga
     4621 actgagatcc gatctgctgg gcttgctgaa ggtagggggt caagaaccag aggtgtaaag
     4681 tacttgagta attttacttg attactgtac ttaagtatta tttttgggga tttttacttt
     4741 acttgagtac aattaaaaat caatactttt acttttactt aattacattt ttttagaaaa
     4801 aaaagtactt tttactcctt acaattttat ttacagtcaa aaagtactta ttttttggag
     4861 atcacttcat tctattttcc cttgctatta ccaaaccaat tgaattgcgc tgatgcccag
     4921 tttaatttaa atgttattta ttctgcctat gaaaatcgtt ttcacattat atgaaattgg
     4981 tcagacatgt tcattggtcc tttggaagtg acgtcatgtc acatctatta ccacaatgca
     5041 cagcaccttg acctggaaat tagggaaatt ataacagtca atcagtggaa gaaaatggag
     5101 gaagtatgtg attcatcagc agctgcgagc agcacagtcc aaaatcagcc acaggatcaa
     5161 gagcacccgt ggccgtatct tcgccccggg agcttcgaat tctaatatgg caataaacga
     5221 tcttcagaga aacttgcatt ggtaaatagt ttcagcctat gcatatgtag ttattcaaac
     5281 tgtaatgact atcttacagt attatatgtt tataatagtt tgatccaata aatccagatt
     5341 tatatgcttt tcaacataaa attgtgaaat gcacagaatg tacatataca atacatttcc
     5401 tacaataatg ctctcttttc tattaatagt gtgcattcag tgcaggatgt tatgtattta
     5461 ttttctgtag tgcaaacttt ttcaaagaat tttccagtct aagcttttca gtcaagaaaa
     5521 aaaaaagcga ttgattatct gtgatattta aaaaaattag ctcttaaatc tggtgaacca
     5581 agtggccaca acacattaaa tactgacgtt ctacagccat gtagcttcat ctggtctggt
     5641 ttgttttggc aggcacttat tttccggtga tagttggcca caagagtatt gtctgagcca
     5701 ttcagtgcta gactgtcatt ctcaggtcag agtccacccc gctgattctg ctgacaacac
     5761 tgttcccacc atgagataat gccattccag agagatccat ttgtaagccc ctctttctgc
     5821 agcacaggta tataaccagg ggtctgcctc cactaaggcc ggcacacatc atttggggat
     5881 ctttgtactg tggatccacc ggtcgccacc atggatggtg agcaagggcg aggagctgtt
     5941 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     6001 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     6061 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
     6121 gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     6181 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     6241 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     6301 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     6361 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     6421 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     6481 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     6541 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     6601 gatcactctc ggcatggacg agctgtacaa gtaaggccgg ccgcgactct agatcataat
     6661 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     6721 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     6781 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6841 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa tcgagggggg gcccggtacc
     6901 caattcgccc tatagtgagt ctcgagccaa gtgcctcaat tcgatcaatc aacgattaaa
     6961 aatattgagt ctgcaaaagg acttgatgtg tgggacagct ggccgctgca aaacgctgac
     7021 ggaacagtag ctgaatacaa cggctatcac gttgtgtttg ctcttgcggg aagcccgaaa
     7081 gacgctgatg acacatcaat ctacatgttt tatcaaaagg tcggcgacaa ctcaatcgac
     7141 agctggaaaa acgcgggccg tgtctttact cgaggggggg cccggtaccc aattcgccct
     7201 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     7261 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     7321 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     7381 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7441 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7501 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     7561 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     7621 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7681 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7741 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7801 acgcgaattt taacaaaata ttaacgctta caatttaggt g
//
