LOCUS       sequence-393502-        7961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC targeting cassette for pIndigoBAC-536 with GalK selection marker
            and lens-specific transgenesis marker dsRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7961)
  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 7961)
  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-IndHom-Tol2-FRT-GalK-cryaa-dsRed
FEATURES             Location/Qualifiers
     source          1..7961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    424..445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(460..465,466..483,484..490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    498..514
                     /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     522..538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        559..577
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    977..1098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(988..1007)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1042..1061
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1223..1900)
                     /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(1285..1308)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1483..1502)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1700..1720
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1752..1770
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1897..1906
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(2944..3131,3132..3143)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     protein_bind    complement(3198..3231)
                     /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)"
     promoter        3235..3282
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(4464..4480)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4464..4480)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(4489..4522)
                     /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(4824..4835,4836..4998)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    5558..5579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5594..5599,5600..5617,5618..5624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5632..5648
                     /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     5656..5672
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5709..5727)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5734..5751)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5734..5750)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5743..5765)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5891..6346)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6028..6049)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6240..6259
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6373..6477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6478..6546,6547..7338)
                     /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(6696..6715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7509..7961,1..136)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      541 tgattacgcc aagctcgaaa ttaaccctca ctaaagggaa caaaagctgg taccgggccc
      601 cccctcgagg tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
      661 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
      721 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
      781 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
      841 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
      901 cgatgagtgg cagggcgggg cgtaattttc tcgagactca ctatagggcg aattgggtac
      961 cgggcccccc ctcgattaag atacattgat gagtttggac aaaccacaac tagaatgcag
     1021 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1081 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1141 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatatggtat ggctgattat
     1201 gatctagagt cgcggccggc cgctacagga acaggtggtg gcggccctcg gcgcgctcgt
     1261 actgctccac gatggtgtag tcctcgttgt gggaggtgat gtccagcttg gagtccacgt
     1321 agtagtagcc gggcagctgc acgggcttct tggccatgta gatggacttg aactccacca
     1381 ggtagtggcc gccgtccttc agcttcaggg ccttgtggat ctcgcccttc agcacgccgt
     1441 cgcgggggta caggcgctcg gtggaggcct cccagcccat agtcttcttc tgcattacgg
     1501 ggccgtcgga ggggaagttc acgccgatga acttcacctt gtagatgaag gagccgtcct
     1561 gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct
     1621 cccacttgaa gccctcgggg aaggacagct tcttgtagtc ggggatttcg gcggggtgct
     1681 tcacgtacac cttggagccg tactggaact ggggggacag gatgtcccag gcgaagggca
     1741 gggggccgcc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     1801 cgccctcgcc ctcgatctcg aattcgtggc cgttcacgga gccctccatg cgcaccttga
     1861 agcgcatgaa ctccttgatg acgtcctcgg agggggccat ggtggcgacc ggtggatcca
     1921 cagtacaaag atccccaaat gatgtgtgcc ggccttagtg gaggcagacc cctggttata
     1981 tacctgtgct gcagaaagag gggcttacaa atggatctct ctggaatggc attatctcat
     2041 ggtgggaaca gtgttgtcag cagaatcagc ggggtggact ctgacctgag aatgacagtc
     2101 tagcactgaa tggctcagac aatactcttg tggccaacta tcaccggaaa ataagtgcct
     2161 gccaaagcaa accagaccag atgaagctac atggctgtag aacgtcagta tttaatgtgt
     2221 tgtggccact tggttcacca gatttaagag ctaatttttt taaatatcac agataatcaa
     2281 tcgctttttt ttttcttgac tgaaaagctt agactggaaa attctttgaa aaagtttgca
     2341 ctacagaaaa taaatacata acatcctgca ctgaatgcac actattaata gaaaagagag
     2401 cattattgta ggaaatgtat tgtatatgta cattctgtgc atttcacaat tttatgttga
     2461 aaagcatata aatctggatt tattggatca aactattata aacatataat actgtaagat
     2521 agtcattaca gtttgaataa ctacatatgc ataggctgaa actatttacc aatgcaagtt
     2581 tctctgaaga tcgtttattg ccatattaga attcgaagct cccggggcga agatacggcc
     2641 acgggtgctc ttgatcctgt ggctgatttt ggactgtgct gctcgcagct gctgatgaat
     2701 cacatacttc ctccattttc ttccactgat tgactgatat aatttcccta atttccaggt
     2761 caaggtgctg tgcattgtgg taatagatgt gacatgacgt cacttccaaa ggaccaatga
     2821 acatgtctga ccaatttcat ataatgtgaa aacgattttc ataggcagaa taaataacat
     2881 ttaaattaaa ctgggcatca gcgcaattca attggtttgg taatagcaag ggaaaataga
     2941 atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt aaggagtaaa
     3001 aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg atttttaatt
     3061 gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat caagtaaaat
     3121 tactcaagta ctttacacct ctggttcttg accccctacc ttcagcaagc ccagcagatc
     3181 ggatctcagt tcggtacgaa gttcctatac tttctagaga ataggaactt ccctgttgac
     3241 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag ggactaaacc
     3301 caggaggcag atcatgagtc tgaaagaaaa aacacaatct ctgtttgcca acgcatttgg
     3361 ctaccctgcc actcacacca ttcaggcgcc tggccgcgtg aatttgattg gtgaacacac
     3421 cgactacaac gacggtttcg ttctgccctg cgcgattgat tatcaaaccg tgatcagttg
     3481 tgcaccacgc gatgaccgta aagttcgcgt gatggcagcc gattatgaaa atcagctcga
     3541 cgagttttcc ctcgatgcgc ccattgtcgc acatgaaaac tatcaatggg ctaactacgt
     3601 tcgtggcgtg gtgaaacatc tgcaactgcg taacaacagc ttcggcggcg tggacatggt
     3661 gatcagcggc aatgtgccgc agggtgccgg gttaagttct tccgcttcac tggaagtcgc
     3721 ggtcggaacc gtattgcagc agctttatca tctcccgctg gacggcgcac aaatcgcgct
     3781 taacggtcag gaagcagaaa accagtttgt aggctgtaac tgcgggatca tggatcagct
     3841 aatttccgcg ctcggcaaga aagatcatgc cttgctgatc gattgccgct cactggggac
     3901 caaagcagtt tccatgccca aaggtgtggc tgtcgtcatc atcaacagta acttcaaacg
     3961 taccctggtt ggcagcgaat acaacacccg tcgtgaacag tgcgaaaccg gtgcgcgttt
     4021 cttccagcag ccagccctgc gtgatgtcac cattgaagag ttcaacgctg ttgcgcatga
     4081 actggacccg atcgtggcaa aacgcgtgcg tcatatactg actgaaaacg cccgcaccgt
     4141 tgaagctgcc agcgcgctgg agcaaggcga cctgaaacgt atgggcgagt tgatggcgga
     4201 gtctcatgcc tctatgcgcg atgatttcga aatcaccgtg ccgcaaattg acactctggt
     4261 agaagtcgtc aaagctgtga ttggcgacaa aggtggcgta cgcatgaccg gcggcggatt
     4321 tggcggctgt atcgtcgcgc tgatcccgga agagctggtg cctgccgtac agcaagctgt
     4381 cgctgaacaa tatgaagcaa aaacaggtat taaagagact ttttacgttt gtaaaccatc
     4441 acaaggagca ggacagtgct gaggatccac tagttctaga gcggccgcga agttcctata
     4501 ctttctagag aataggaact tctaactaac caagatccgc tggctaagaa ctcatcagcc
     4561 tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg gctactacat
     4621 ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca atgctcttgg
     4681 atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa tgccctctga
     4741 tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc acacaggtca
     4801 gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt
     4861 gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa
     4921 tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attaaggaag
     4981 taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa tctcaaaaaa
     5041 cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta tttaattatc
     5101 aaactataag acagacaatc taacgccagt acacgctact caaagttgta aaacctcaga
     5161 tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc tcttttgggg
     5221 ctgaaaagca atcccgcagt gctgaaaagc ctctcacagg cagccgatgc gggaagaggt
     5281 gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga ctccctggtg
     5341 tctgaaacaa ggccagatgc ggccgccacc gcggggttgc tacgcctgaa taagtgataa
     5401 taagcggatg aatggcagaa attcgatgat aagctgtcaa acatgagaat tggtcgacgg
     5461 cccgggcggc cgcaaggggt tcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5521 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     5581 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     5641 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcta tttagagctc
     5701 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     5761 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5821 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5881 tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5941 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6001 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6061 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6121 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6181 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6241 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6301 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     6361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6661 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6721 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7381 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7621 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7861 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7921 cccgaaggga gaaaggcgga caggtatccg gtaagcggca g
//
