LOCUS       sequence-344912-        3609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pME-Vector with codon improved iCre sequence. Tol2-kit compatible. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3609)
  AUTHORS   Tromp A, Wang H, Hall TE, Mowry B, Giacomotto J
  TITLE     Optimising the zebrafish Cre/Lox toolbox. Codon improved iCre, new 
            gateway tools, Cre protein and guidelines.
  JOURNAL   Front Physiol. 2023 Aug 2;14:1221310. doi: 
            10.3389/fphys.2023.1221310. eCollection 2023.
  PUBMED    37601640
REFERENCE   2  (bases 1 to 3609)
  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: "Front 
            Physiol."; date: "2023-08-2"; volume: "14:1221310. doi"; pages: " 
            10.3389/fphys.2023.1221310. eCollection 2023"
COMMENT     Sequence Label: 469_pME_iCre
FEATURES             Location/Qualifiers
     source          1..3609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(154..742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(234..253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(889..1698)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1612..1631
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1690..1709
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1811..1827
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1811..1827
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1832..1851
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1832..1850
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1868..1967
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2998..3018)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3021..3030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3033..3132)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3148..3165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3148..3164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3157..3179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3228..3314
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3406..3433
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3592..3609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
       61 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      781 gtggaacgac gcgtaactca cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca
      841 agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc
      901 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      961 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     1021 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     1081 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     1141 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     1201 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     1261 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     1321 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     1381 ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     1441 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     1501 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     1561 atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     1621 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc
     1681 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     1741 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg
     1801 ccagagctgc caggaaacag ctatgaccat gtaatacgac tcactatagg ggatatcagc
     1861 tggatggcaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     1921 taagcaatgc tttcttataa tgccaacttt gtacaagaaa gctgggtgtc agtccccatc
     1981 ctcgagcagc ctcaccatgg ccccagtctc agagtccagg tttctgatgt agttcatcac
     2041 aatgttcaca ttggtccagc caccagcctg catgatttca gggatggaca caccagccct
     2101 ggccatgtcc ctggcagcac ccactctggc agagtggcca gaccaggcca ggtatctctg
     2161 cccagagtca tccttggcac catagatcag gcggtgggtg gcctcaaaga tcccttccag
     2221 ggcccgggtg gacagttggg aggtggcaga aggggcagcc acaccattct ttctgacccg
     2281 gcagaacagg tagttgttgg ggtcatcagc cacaccagac acagagatcc atctctccac
     2341 cagcttggta acccccaggg acagggcctt ctccacacca gctgtggaca ccagggtctt
     2401 ggtcctgcca atgtggatca gcattctccc accatcggtg cgggagatgt ccttcactct
     2461 gattctggca atttcggcaa tgcgcagcag ggtgttgtag gcaatgccca ggaaggccag
     2521 gttcctgatg tcctggcatc tgtcagagtt ctccatcagg gatctgactt ggtcaaagtc
     2581 agtgcgttca aaggccaggg cctgcttggc tctctcccca gcatccacat tctcctttct
     2641 gattctcctc atcaccaggg acacagcatt ggagtcagaa gggcgaggca ggccagatct
     2701 cctgtgcagc atgttgagct ggcccaggtg ctgttggatg gtcttcacag ccaggcctct
     2761 ggcttgcagg tacaggaggt agtccctcac atcctcaggt tcagcaggga accatttcct
     2821 gttgttcagc ttgcaccagg cagcccagga tctgcacaca gacaggagca tcttccaggt
     2881 gtgttcagag aaggcctgcc tgtccctgaa catgtccatc aggttcttcc tgacttcatc
     2941 agaggtggca tccacaggga gggcaggcag gttttggtgc acagtcagca ggttggagac
     3001 tttcctcttc ttcttgggca ccatggtggc ggagcctgct tttttgtaca aagttggcat
     3061 tataaaaaag cattgctcat caatttgttg caacgaacag gtcactatca gtcaaaataa
     3121 aatcattatt tggggcccga gcttaagact ggccgtcgtt ttacaacgtc gtgactggga
     3181 aaacatccat gctagcgtta acgcgagagt agggaactgc caggcatcaa ataaaacgaa
     3241 aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     3301 tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt
     3361 ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga
     3421 cggatggcct ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc
     3481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3601 tgactcgct
//
