LOCUS       sequence-254265-        6554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tol2 bicistronic vector expressing PcyA and HO1 for Phycocyanobilin 
            (PCB) synthesis, driven by heatshock promoter hsp70..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6554)
  AUTHORS   Yen ST, Trimmer KA, Aboul-Fettouh N, Mullen RD, Culver JC, Dickinson
            ME, Behringer RR, Eisenhoffer GT
  TITLE     CreLite: An optogenetically controlled Cre/loxP system using red 
            light.
  JOURNAL   Dev Dyn. 2020 Nov;249(11):1394-1403. doi: 10.1002/dvdy.232. Epub 
            2020 Aug 31.
  PUBMED    32745301
REFERENCE   2  (bases 1 to 6554)
  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"; doi: "10.1002/dvdy.232"; 
            journalName: "Dev Dyn"; date: "2020-11"; volume: "249"; issue: "11";
            pages: "1394-1403"
COMMENT     Sequence Label: pTol2-PcyA-IRES-HO1
FEATURES             Location/Qualifiers
     source          1..6554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..12,13..200)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     protein_bind    272..292
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1809..1833
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1840..1914
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     protein_bind    complement(2650..2674)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(2675..3249,3250..3252,3253..3261)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2842..2859)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3069..3088
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3262..3336
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     protein_bind    complement(4078..4098)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    4126..4247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4137..4156)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4191..4210
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(4373..4535,4536..4547)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4564..4581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4735..5323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4815..4834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5494..6285,6286..6354))
                     /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     6117..6136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6355..6459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6527..6545
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cagaggtgta aagtacttga gtaattttac ttgattactg tacttaagta ttatttttgg
       61 ggatttttac tttacttgag tacaattaaa aatcaatact tttactttta cttaattaca
      121 tttttttaga aaaaaaagta ctttttactc cttacaattt tatttacagt caaaaagtac
      181 ttattttttg gagatcactt cattctattt tcccttgcta ttaccaaacc aattgaattg
      241 cgctgatgcc cagtttaatt taaatagatc tcaactttgt atagaaaagt tgtcaggggt
      301 gtcgcttggt tatttccaaa aatcaaatta attttattaa actattagaa cgagcatgtt
      361 ttgtctatat gctacagaag ataaaaaata ataggagtta acagttataa aacaacacac
      421 tttgtttcta ttgattgttg accacactgg ggtctcatta agttagatta aagacacact
      481 aactgggtca aaagcagcag attgatttca tagcaccagg gtaaactttc taacactttt
      541 acggcaatca tatacattaa aattaaatac agaccacgac tgaacaagga ggatgatctc
      601 caatattaaa caaagagact tgtgcctatt tctctgaggg taaacatgac ctctcaagtt
      661 agcaagttgg ttttaacact acaaaaatag ttaagactgc aatcccagaa taaagtattg
      721 gttttaacca atcaatatag tacagtaaac atccatttgt tttgttgaaa cgttaaacaa
      781 atctgaccaa agctattagc ttatataaaa caggtttgcc ttctatgtag ctgaaaacac
      841 cacaggcccg attttgctac tgtgtaaaac atttcagcaa gattttttta ttgcattttt
      901 ttttactgaa tcgttcaaac attttatcat tttagtttgt tcattcattg caactggaaa
      961 aacaacacat cacacaaccg cacatatttc agcaataagt acaataaaac actcaaataa
     1021 aaaaaacatt ttaaatctct ttgtattttt gaccgctgtt tcgcgtaatt tcacggtaaa
     1081 actctggaaa tctccactac attcctctca gcggctcctc tcaatgacag ctgaagaagt
     1141 gacgcggctg cctgctgtgt tttgattggt cgaattcact ggaggcttcc agaacagtgt
     1201 agagtctgaa cgggtgcgcg ctctgctgta tttaaagggc gaaagagaga ccgcagagaa
     1261 actcaaccga agagaagcga cttgacaaag aagaaaagag cagcctgaca ggactttttc
     1321 cccgacgagg tgtttattcg ctctatttaa gaatctactg taaggtaagt ctcaatatat
     1381 tgtactctat tggctaatca gaattatata gagattatat gtacttaatg tcaaaaaatc
     1441 aactttgtat atgtaatctt tttacatgtg gactgcctat gttcatctta ttttaggtct
     1501 actagaaaat tatatttccc gttttcacaa taaggatttt taaaaaaagc aatgaacaga
     1561 cgggcattta ctttatgttg ctgacattat tttatatgag cataataacc ataaatacta
     1621 gcaaatgtcc taaatgaatt tgtgttaatg ttgtctacaa aagaaaatta gcgttttact
     1681 tgtacaacta ataataactt ggttattaag agaatttcac ttgttgacta gaaaaatcct
     1741 ttcataatga aacaattgca ccataaattg tataaatata aaattaattc taattgtttt
     1801 tttttcctgc aagtttgtac aaaaaagcag gctgccacca tgtccgtcct gacgccgctg
     1861 ctgctgcggg gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat
     1921 tcgttgggta ccgccaccat gtctctgaga cagcaccagc accccctgat ccagagactg
     1981 gccgacagaa tcgaggccat ctggcaggcc tttttccccc tggcccctta cgccctgcct
     2041 gaggatctgg gctacgtgga aggcaagctg gaaggcgagc ggctgaccat cgagaaccac
     2101 tgctaccagg cccctccatt ccggaagctg cacctggaac tggccagagt gggagagtcc
     2161 ctggacatcc tgcactgcgt gatgttcccc gagcccagat acgacctgcc catgttcggc
     2221 tgcgacctcg tgggaggcag aggccagatc tctgccgcca tcgtggatct gtctcccgtg
     2281 accggacagc tgcctgccgc ttacacctgt gccctgaatg ccctgcccaa gctgaccttc
     2341 cggcagccca gagaactgcc tccttggggc cacatcttca gccctttctg catcttcatc
     2401 cggccccagg gcgaggccga ggaacagcag ttcctggatc ggatcggcga gtacctgacc
     2461 ctgcactgtc agctgtccca gcaggccgtg cctaccgatc atccacaggc tgtgatcgcc
     2521 ggccagagac agtactgcca acagcagcag cagaacgaca agaccagacg ggtgctggaa
     2581 aaggccttcg gcgtgccatg ggccgagcgg tatatgacca ccgtgctgtt cgacgtgccc
     2641 cccgtgtgaa cccagctttc ttgtacaaag tgggcccctc tccctccccc ccccctaacg
     2701 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     2761 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2821 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2881 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2941 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3001 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3061 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3121 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3181 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3241 gatgataata tggccacaac catgtccgtc ctgacgccgc tgctgctgcg gggcttgaca
     3301 ggctcggccc ggcggctccc agtgccgcgc gccaagatcc attcgttggg taccgccacc
     3361 atgacaacat ctctggccac caagctgaga gagggcacca agaaagccca caccatggcc
     3421 gagaacgtgg gcttcgtgcg gtgcttcctg aagggaaccg tggaaaagtc ctcctaccgg
     3481 aagctggtgg cctccctgta ccacgtgtac tccgccatgg aacaggaaat ggaacggctg
     3541 aaggaccacc ccatcgtggg caagatctac ttccccgagc tgaaccggaa gtcctccctg
     3601 gaacgggacc tgacctacta cttcggctcc aactggcggg aagagatccc cccatctcct
     3661 gccacccagg cctacgtggc cagaatccac gaggtggcca ataccgcccc tgagctgctg
     3721 gtggctcact cctacaccag atacctgggc gacctgtctg gcggccagat tctgaagggc
     3781 attgccgagc gggccatgaa cctgcaggat ggcgagggca ccgccttcta cagattcgag
     3841 tccatctccg acgagaaggc ctttaagcag ctgtaccggc agcggctgga cgagctgcct
     3901 gtggatgagg ccaccgccga cagaatcgtg gacgaggcca atgccgcctt cggcatgaac
     3961 atgaagatct ttcaggaact ggaaggcaac ctgatccggg ccatcggcca gctgctgttc
     4021 aacaccctga cccggcggaa gcagagaggc tctaccgagc tggccaccgc tgactgacaa
     4081 ctttattata catagttgat ggccggccgc ttcgagcaga catgataaga tacattgatg
     4141 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4201 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4261 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     4321 acctctacaa atgtggtaga tatcaagctt aaacaagaat ctctagtttt ctttcttgct
     4381 tttactttta cttccttaat actcaagtac aattttaatg gagtactttt ttacttttac
     4441 tcaagtaaga ttctagccag atacttttac ttttaattga gtaaaatttt ccctaagtac
     4501 ttgtactttc acttgagtaa aatttttgag tactttttac acctctgggc gctcttccgc
     4561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     5641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6481 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6541 gaggcccttt cgtc
//
