LOCUS       sequence-258340-        6031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for production of codon optimized Tol2 
            transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6031)
  AUTHORS   Mackey AS, Redd PS, DeLaurier A, Hancock CN
  TITLE     Codon optimized Tol2 transposase results in increased transient 
            expression of a crystallin-GFP transgene in zebrafish.
  JOURNAL   MicroPubl Biol. 2020 Jun 11;2020:10.17912/micropub.biology.000268. 
            doi: 10.17912/micropub.biology.000268.
  PUBMED    32626847
REFERENCE   2  (bases 1 to 6031)
  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: "MicroPubl 
            Biol."; date: "2020-06-11"; volume: 
            "2020:10.17912/micropub.biology.000268. doi"; pages: " 
            10.17912/micropub.biology.000268"
COMMENT     Sequence Label: pCS2FA CO Tol2 TPase
FEATURES             Location/Qualifiers
     source          1..6031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..1031
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1066..1084
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1066..1083
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1121..1130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1127..3076
                     /codon_start=1
                     /product="transposase of the Tol2 transposon from the 
                     medaka fish Oryzias latipes (Kawakami, 2007)"
                     /label=Tol2 transposase
                     /translation="MEEVCDSSAAASSTVQNQPQDQEHPWPYLREFFSLSGVNKDSFKM
                     KCVLCLPLNKEISAFKSSPSNLRKHIERMHPNYLKNYSKLTAQKRKIGTSTHASSSKQL
                     KVDSVFPVKHVSPVTVNKAILRYIIQGLHPFSTVDLPSFKELISTLQPGISVITRPTLR
                     SKIAEAALIMKQKVTAAMSEVEWIATTTDCWTARRKSFIGVTAHWINPGSLERHSAALA
                     CKRLMGSHTFEVLASAMNDIHSEYEIRDKVVCTTTDSGSNFMKAFRVFGVENNDIETEA
                     RRCESDDTDSEGCGEGSDGVEFQDASRVLDQDDGFEFQLPKHQKCACHLLNLVSSVDAQ
                     KALSNEHYKKLYRSVFGKCQALWNKSSRSALAAEAVESESRLQLLRPNQTRWNSTFMAV
                     DRILQICKEAGEGALRNICTSLEVPMFNPAEMLFLTEWANTMRPVAKVLDILQAETNTQ
                     LGWLLPSVHQLSLKLQRLHHSLRYCDPLVDALQQGIQTRFKHMFEDPEIIAAAILLPKF
                     RTSWTNDETIIKRGMDYIRVHLEPLDHKKELANSSSDDEDFFASLKPTTHEASKELDGY
                     LACVSDTRESLLTFPAICSLSIKTNTPLPASAACERLFSTAGLLFSPKRARLDTNNFEN
                     QLLLKLNLRFYNFE"
     polyA_signal    3114..3248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3138..3157)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3192..3211
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3361..3381)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3361..3379)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3400..3416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3400..3416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3413..3435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3424..3440
                     /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(3448..3454,3455..3472,3473..3478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3493..3514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3631..3648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3802..4390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3882..3901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4561..5352,5353..5421))
                     /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     5184..5203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5422..5526)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5552..6007)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5689..5710)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5901..5920
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
       61 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
      121 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
      181 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
      241 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
      301 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
      361 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
      421 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
      481 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
      541 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
      601 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
      661 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
      721 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
      781 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
      841 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
      901 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
      961 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     1021 gtttagggaa ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta
     1081 tagaatacaa gctacttgtt ctttttgcag gatccggccg gccaccatgg aagaggtatg
     1141 tgacagtagt gcggcggctt cttctactgt tcaaaaccaa cctcaggatc aggaacatcc
     1201 ctggccatat cttcgagaat tcttttccct gtctggcgtt aataaggata gctttaagat
     1261 gaagtgcgtg ttgtgccttc cgttgaacaa ggaaatatcc gcattcaagt cttctcctag
     1321 taatctccga aagcacatag agaggatgca cccaaattat ctgaagaatt attcaaaatt
     1381 gacagcacag aagaggaaaa taggaactag cactcatgca agcagttcaa aacaacttaa
     1441 ggtcgattct gtgtttccag taaagcatgt tagtccagtc acagtaaaca aagcaatcct
     1501 tcgatacata attcaaggtc tgcatccatt ctcaacggtg gatcttccca gcttcaagga
     1561 acttatttcc acgctgcagc ccggcatttc agtaatcacc aggccaacat tgcggtccaa
     1621 gatcgccgag gctgcgctga ttatgaaaca gaaggttact gccgcgatgt cagaagtcga
     1681 gtggatagct actactaccg actgctggac tgcaaggcga aaaagtttca ttggcgtgac
     1741 agcacactgg atcaatcctg gtagcctcga gaggcactcc gctgccctgg cttgtaaacg
     1801 ccttatgggg agtcatactt ttgaagtcct tgcatctgca atgaatgata tccacagtga
     1861 gtatgagatt cgtgacaagg ttgtttgtac tacaacagat tcaggtagca actttatgaa
     1921 agctttccgt gtattcggag ttgagaacaa tgatatcgaa acagaagcca gaaggtgcga
     1981 gagtgacgac acggactccg aagggtgtgg tgagggttcc gatggcgttg aatttcagga
     2041 cgcgtctagg gttcttgacc aagatgacgg ctttgaattt cagttgccga aacatcagaa
     2101 atgtgcgtgc catctgctca atcttgtaag ttcagtcgac gcccagaaag ctctttctaa
     2161 cgaacactat aagaaactct atagaagcgt cttcgggaag tgtcaagcgc tctggaacaa
     2221 gtccagccgt tctgcacttg cggcagaggc tgtagaatcc gaaagtcgcc tgcaactgtt
     2281 gcgacccaat caaacgcgat ggaactcaac cttcatggcc gtggaccgga ttctccagat
     2341 ctgcaaggaa gcgggcgagg gtgccctccg caatatttgt acttctttgg aggtaccaat
     2401 gtttaatccg gcagaaatgc ttttcttgac ggaatgggcc aatacaatgc gtccggtggc
     2461 aaaggttctt gacattctgc aggcggaaac aaatactcaa ctcggttggc tgctgccgag
     2521 cgtacaccaa ctctccctga agctccagcg tctccatcat agtttgcggt actgcgaccc
     2581 gctggtagat gcgctccagc aagggattca gactcgcttt aaacatatgt tcgaagatcc
     2641 agagataatc gctgcggcga tactgcttcc caaattcaga acaagctgga ctaatgacga
     2701 gaccatcatc aagcggggga tggattacat acgggtacac cttgaaccac tcgaccataa
     2761 gaaagaactg gcaaactctt cttccgatga tgaggacttc ttcgcttcac ttaaaccaac
     2821 gacgcatgaa gcgtcaaagg agctcgacgg ttatctcgcg tgcgtttcag acaccagaga
     2881 atccctcttg acctttcccg cgatatgcag cttgtccata aagaccaata cgccactgcc
     2941 tgctagcgct gcctgtgaac gtctcttttc cactgccggg ttgctcttta gcccaaagag
     3001 agccaggctt gacacgaaca actttgagaa ccaacttctc ttgaagctca accttaggtt
     3061 ttacaacttc gagtagggcg cgcctctaga actatagtga gtcgtattac gtagatccag
     3121 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3181 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3241 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     3301 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     3361 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3421 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     3481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5581 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5641 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5701 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5761 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5821 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5881 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5941 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6001 ggcgcgtccc attcgccatt caggctgcgc a
//
