LOCUS       sequence-135629-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tol2 vector with multi cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Villefranc JA, Amigo J, Lawson ND
  TITLE     Gateway compatible vectors for analysis of gene function in the 
            zebrafish.
  JOURNAL   Dev Dyn. 2007 Nov . 236(11):3077-87.
  PUBMED    17948311
REFERENCE   2  (bases 1 to 5863)
  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: "Dev Dyn. 
            2007 Nov . 236(11):3077-87."
COMMENT     Sequence Label: 240 pTol2005b
FEATURES             Location/Qualifiers
     source          1..5863
                     /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)"
     misc_feature    join(3393..3580,3581..3592)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3626..3642)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3626..3642)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3679..3699)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3679..3697)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3718..3734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3718..3734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3731..3753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3742..3758
                     /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(3766..3772,3773..3790,3791..3796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3811..3832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3949..3966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4120..4708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4200..4219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4879..5670,5671..5739))
                     /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     5502..5521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5740..5844)
                     /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 tatcgctagc atcgatgcat gggccctcga gcatatgaga tctgtcccga tctttctctt
     1381 ctgtgctgtc aatttagagt agtttttgag gtaatttggg tgcattctct gtaaaaacaa
     1441 caaataaata tttatattgt atgcaataaa tgtaacaatt acacatacac tctctgatta
     1501 cagcattaaa gggttagttc acccaaaaat gaaaagatgt cattaatgac tcaccctcat
     1561 gtcgttccaa gcccgtaaca cctccgttca tcttcggaac acagtttaag atattttaga
     1621 tttagtccga gagctttctg tccctccact gaaaatgtat gtacggtata ctgtccatgt
     1681 ccagaaaggt aataaaaaca tcaaagtagt ccatgtgaca tcagtgggtt aggtagaatt
     1741 atttgaagca tcgaaaatac attttggtcc aaaaataaca aaacctacga ctttattcag
     1801 cattgtagcg tcactgcgaa gtcgtgaacg cggattgaca acagacccgg aagagaatac
     1861 aatgccgaat aaagtcgtag gttttgttat ttttggacca aaatgtattc gatgcttcaa
     1921 aaaattctaa ctaacccact gatgtcacat ggactacttt gatgttttta ttacctttct
     1981 ggacatggac agtataccgt acatacattc tcaatggagg cacagaaagc tctcggacta
     2041 aatctaaaat atcttaaact gtgttctgaa gatgaacgga ggtcttacgg gcttggaacg
     2101 acatgagggc gagtcattaa tgacattatt ttcatttttg ggtgaactaa ccctttaata
     2161 aaactgacat ttgcagccaa ttgcacactt tataatgtta aatacttgtt gattttagaa
     2221 atacagtagc atttagtgac aatcaaacat taattttatt tctgattata ttgacaaaaa
     2281 cacaggactg taaggctgac atgacagggt gaattgattt cattacatta gttttgatgg
     2341 taacactttt acaataaggt tcgttagtta actacattag ttaacatgaa ctaataatga
     2401 actgcacgta tacatgttaa tttcaacatt tactaatact ttatatcttg ttaacattag
     2461 ttaatgcact gtgaactaac atgaacaaac aatgaacatc tgtatttcta ttaactaacg
     2521 ttaacaaaga ttaacagata cagtaacaaa tgtattgctc atggttattt aatgttagtt
     2581 aatacatgaa ctaatcaacc tcattgtaaa gtgttaccgt tttgacaata caacatgttt
     2641 ggaaaaaaat ttctcgcttg gttttcttgt gttttgcatg aattcaaaca caatgagagc
     2701 acggaatgat aatgattatt caaagctgca ataggcgcca aatttcccgt ggacctaacg
     2761 ttaccaatta caaatgcatt ttagcatttt attacgtgag ccttatattg tcaatcgcat
     2821 gcattagtga aaataggcct actacacata taaaaggcgc gccatcaacg tcaaaacatg
     2881 cacacccaaa aaaaaaaaaa aaaaaaaaaa aaaaaactat cagtaaaaca aaatacttaa
     2941 tgtacttacc tcaatatgct tccttaggtt tgatggcgaa cttttgaagg ccgatatttc
     3001 tttattaagc gggagacaga ggacacattt catcttgaat gaatctttat ttacaccact
     3061 taaagaaaag aattcgcgaa gatacggcca cgggtgctct tgatcctgtg gctgattttg
     3121 gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct tccactgatt
     3181 gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt aatagatgtg
     3241 acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata taatgtgaaa
     3301 acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag cgcaattcaa
     3361 ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt
     3421 tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta
     3481 agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat
     3541 aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc tggttcttga
     3601 ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc caccgcggtg
     3661 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4561 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtg
//
