LOCUS       sequence-203031-        8569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heat shock inducible transgenesis, Galactsyl transferase 
            (trans-golgi marker).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8569)
  AUTHORS   Icha J, Kunath C, Rocha-Martins M, Norden C
  TITLE     Independent modes of ganglion cell translocation ensure correct 
            lamination of the zebrafish retina.
  JOURNAL   J Cell Biol. 2016 Oct 24;215(2):259-275.
  PUBMED    27810916
REFERENCE   2  (bases 1 to 8569)
  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: "J Cell 
            Biol."; date: "2016-10-24"; volume: "215(2)"; pages: "259-275"
COMMENT     Sequence Label: hsp70:GalT-RFP
FEATURES             Location/Qualifiers
     source          1..8569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(778..789,790..952)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    1324..1344
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    2864..2888
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3107..3116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3113..3790
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3113..3790
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3243..3261)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3511..3530
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3705..3728
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(3793..3817)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3822..3956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3846..3865)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3900..3919
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(6099..6286,6287..6298)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6332..6348)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6332..6348)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6385..6405)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6385..6403)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6424..6440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6424..6440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6437..6459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6448..6464
                     /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(6472..6478,6479..6496,6497..6502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6517..6538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6655..6672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6826..7414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6906..6925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7585..8376,8377..8445))
                     /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     8208..8227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8446..8550)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 caggtcattg cacaacacca gaaatgccct ctgatctgca aaagacgtga atatctgttc
      721 agacacccat atccactctg ttccacacag gtcagaggtt tgtccaggag ttcttgacag
      781 aggtgtaaaa agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt
      841 ttactcaatt aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt
      901 actccattaa aattgtactt gagtattaag gaagtaaaag taaaagcaag aaagaaaact
      961 agagattctt gtttaagctt ttaatctcaa aaaacattaa atgaaatgca tacaaggttt
     1021 tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac aatctaatgc
     1081 cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa gctgattctc
     1141 aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg cagtgctgaa
     1201 aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag agaggctgca
     1261 aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca gatgggccga
     1321 tatcaacttt gtatagaaaa gttgcttcag gggtgtcgct tggttatttc caaaaatcaa
     1381 attaatttta ttaaactatt agaacgagca tgttttgtct atatgctaca gaagataaaa
     1441 aataatagga gttaacagtt ataaaacaac acactttgtt tctattgatt gttgaccaca
     1501 ctggggtctc attaagttag attaaagaca cactaactgg gtcaaaagca gcagattgat
     1561 ttcatagcac cagggtaaac tttctaacac ttttacggca atcatataca ttaaaattaa
     1621 atacagacca cgactgaaca aggaggatga tctccaatat taaacaaaga gacttgtgcc
     1681 tatttctctg agggtaaaca tgacctctca agttagcaag ttggttttaa cactacaaaa
     1741 atagttaaga ctgcaatccc agaataaagt attggtttta accaatcaat atagtacagt
     1801 aaacatccat ttgttttgtt gaaacgttaa acaaatctga ccaaagctat tagcttatat
     1861 aaaacaggtt tgccttctat gtagctgaaa acaccacagg cccgattttg ctactgtgta
     1921 aaacatttca gcaagatttt tttattgcat ttttttttac tgaatcgttc aaacatttta
     1981 tcattttagt ttgttcattc attgcaactg gaaaaacaac acatcacaca accgcacata
     2041 tttcagcaat aagtacaata aaacactcaa ataaaaaaaa cattttaaat ctctttgtat
     2101 ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg gaaatctcca ctacattcct
     2161 ctcagcggct cctctcaatg acagctgaag aagtgacgcg gctgcctgct gtgttttgat
     2221 tggtcgaatt cactggaggc ttccagaaca gtgtagagtc tgaacgggtg cgcgctctgc
     2281 tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa ccgaagagaa gcgacttgac
     2341 aaagaagaaa agagcagcct gacaggactt tttccccgac gaggtgttta ttcgctctat
     2401 ttaagaatct actgtaaggt aagtctcaat atattgtact ctattggcta atcagaatta
     2461 tatagagatt atatgtactt aatgtcaaaa aatcaacttt gtatatgtaa tctttttaca
     2521 tgtggactgc ctatgttcat cttattttag gtctactaga aaattatatt tcccgttttc
     2581 acaataagga tttttaaaaa aagcaatgaa cagacgggca tttactttat gttgctgaca
     2641 ttattttata tgagcataat aaccataaat actagcaaat gtcctaaatg aatttgtgtt
     2701 aatgttgtct acaaaagaaa attagcgttt tacttgtaca actaataata acttggttat
     2761 taagagaatt tcacttgttg actagaaaaa tcctttcata atgaaacaat tgcaccataa
     2821 attgtataaa tataaaatta attctaattg tttttttttc ctgccaagtt tgtacaaaaa
     2881 agcaggctgg atgaggcttc gggagccgct cctgagcggc agcgccgcga tgccaggcgc
     2941 gtccctacag cgggcctgcc gcctgctcgt ggccgtctgc gctctgcacc ttggcgtcac
     3001 cctcgtttac tacctggctg gccgcgacct gagccgcctg ccccaactgg tcggagtctc
     3061 cacaccgctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggcctc
     3121 ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa
     3181 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     3241 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     3301 tcagttccag tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     3361 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     3421 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     3481 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     3541 ggaggcctcc accgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagat
     3601 gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacatggc
     3661 caagaagccc gtgcagctgc ccggcgccta caagaccgac atcaagctgg acatcacctc
     3721 ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac
     3781 cggcgcctaa agacccagct ttcttgtaca aagttgatgt agatccagac atgataagat
     3841 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3901 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3961 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     4021 tcgcggccgc ggcgccaatg cattgggccc tcgagcatat gagatctgtc ccgatctttc
     4081 tcttctgtgc tgtcaattta gagtagtttt tgaggtaatt tgggtgcatt ctctgtaaaa
     4141 acaacaaata aatatttata ttgtatgcaa taaatgtaac aattacacat acactctctg
     4201 attacagcat taaagggtta gttcacccaa aaatgaaaag atgtcattaa tgactcaccc
     4261 tcatgtcgtt ccaagcccgt aacacctccg ttcatcttcg gaacacagtt taagatattt
     4321 tagatttagt ccgagagctt tctgtccctc cactgaaaat gtatgtacgg tatactgtcc
     4381 atgtccagaa aggtaataaa aacatcaaag tagtccatgt gacatcagtg ggttagttag
     4441 aattatttga agcatcgaaa atacattttg gtccaaaaat aacaaaacct acgactttat
     4501 tcagcattgt agcgtcactg cgaagtcgtg aacgcggatt gacaacagac ccggaagaga
     4561 atacaatgcc gaataaagtc gtaggttttg ttatttttgg accaaaatgt attcgatgct
     4621 tcaaaaaatt ctaactaacc cactgatgtc acatggacta ctttgatgtt tttattacct
     4681 ttctggacat ggacagtata ccgtacatac attctcaatg gaggcacaga aagctctcgg
     4741 actaaatcta aaatatctta aactgtgttc tgaagatgaa cggaggtctt acgggcttgg
     4801 aacgacatga gggcgagtca ttaatgacat tattttcatt tttgggtgaa ctaacccttt
     4861 aataaaactg acatttgcag ccaattgcac actttataat gttaaatact tgttgatttt
     4921 agaaatacag tagcatttag tgacaatcaa acattaattt tatttctgat tatattgaca
     4981 aaaacacagg actgtaaggc tgacatgaca gggtgaattg atttcattac attagttttg
     5041 atggtaacac ttttacaata aggttcgtta gttaactaca ttagttaaca tgaactaata
     5101 atgaactgca cgtatacatg ttaatttcaa catttactaa tactttatat cttgttaaca
     5161 ttagttaatg cactgtgaac taacatgaac aaacaatgaa catctgtatt tctattaact
     5221 aacgttaaca aagattaaca gatacagtaa caaatgtatt gctcatggtt atttaatgtt
     5281 agttaataca tgaactaatc aacctcattg taaagtgtta ccgttttgac aatacaacat
     5341 gtttggaaaa aaatttctcg cttggttttc ttgtgttttg catgaattca aacacaatga
     5401 gagcacggaa tgataatgat tattcaaagc tgcaataggc gccaaatttc ccgtggacct
     5461 aacgttacca attacaaatg cattttagca ttttattacg tgagccttat attgtcaatc
     5521 gcatgcatta gtgaaaatag gcctactaca catataaaag gcgcgccatc aacgtcaaaa
     5581 catgcacacc caaaaaaaaa aaaaaaaaaa aaaaaaaaaa aactatcagt aaaacaaaat
     5641 acttaatgta cttacctcaa tatgcttcct taggtttgat ggcgaacttt tgaaggccga
     5701 tatttcttta ttaagcggga gacagaggac acatttcatc ttgaatgaat ctttatttac
     5761 accacttaaa gaaaagaatt cgcgaagata cggccacggg tgctcttgat cctgtggctg
     5821 attttggact gtgctgctcg cagctgctga tgaatcacat acttcctcca ttttcttcca
     5881 ctgattgact gttataattt ccctaatttc caggtcaagg tgctgtgcat tgtggtaata
     5941 gatgtgacat gacgtcactt ccaaaggacc aatgaacatg tctgaccaat ttcatataat
     6001 gtgaaaacga ttttcatagg cagaataaat aacatttaaa ttaaactggg catcagcgca
     6061 attcaattgg tttggtaata gcaagggaaa atagaatgaa gtgatctcca aaaaataagt
     6121 actttttgac tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg
     6181 taattaagta aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc
     6241 aaaaataata cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggt
     6301 tcttgacccc ctaccttcag caagcccagc agatccacta gttctagagc ggccgccacc
     6361 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     6421 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6481 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6541 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6601 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6661 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6721 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6781 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6841 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6901 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6961 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7021 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7081 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7141 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7201 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7261 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7321 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7381 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7441 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7501 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7561 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7681 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7741 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7921 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7981 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8041 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8101 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8161 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8221 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8281 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8341 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8401 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8461 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8521 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtg
//
