LOCUS       sequence-117433-        5527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of mRNA encoding Tol1 Transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5527)
  AUTHORS   Shin M, Male I, Beane TJ, Villefranc JA, Kok FO, Zhu LJ, Lawson ND
  TITLE     Vegfc acts through ERK to induce sprouting and differentiation of 
            trunk lymphatic progenitors.
  JOURNAL   Development. 2016 Oct 15;143(20):3785-3795. Epub 2016 Sep 12.
  PUBMED    27621059
REFERENCE   2  (bases 1 to 5527)
  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: 
            "Development."; date: "2016-10-15"; volume: "143(20)"; pages: 
            "3785-3795. Epub 2016 Sep 12"
COMMENT     Sequence Label: pToneTP
FEATURES             Location/Qualifiers
     source          1..5527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        435..453
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3053..3187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3077..3096)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3131..3150
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3306..3322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3306..3322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3319..3341)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3330..3346
                     /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(3354..3360,3361..3378,3379..3384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3399..3420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3537..3554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3708..4296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3788..3807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4467..5258,5259..5327))
                     /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     5090..5109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5328..5432)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5500..5518
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tagctaatac gactcactat aggagagaat acaagctact tgttcttttt
      481 gcacatctaa aatatggaga aaaaaaggtc aaagccatct ggtgcccaat ttagaaagaa
      541 aagaaaagaa gaagaggaga aaagagataa agaaaagggg gcacttctaa gatattttgg
      601 atcgtctacc actgctcaag atgagacatc tacctccctg ccagctatct catcagccac
      661 agtcacagtc tcaccccctc aggatgagct accatctaca tcctctgcta ctcatgtagt
      721 tccacagttg ttacctgagc aaagttttga tagtgaggct gaagacgttg ttccatctac
      781 gtctacccag cttgagactt cagaaatgcc tggtgatgaa accccactga ccccgactgc
      841 tgaggaccag cctctaccaa ctgaccctgc aaagtggccc tcacctctga ctgacaggat
      901 acggatggag ctggttcgaa gaggaccaag tagcatacca cctgactttg ttttcccaag
      961 aaatgacagt gatgggagaa gttgtcatca ccactatttc aggaagacac tagtaagtgg
     1021 tgaaaaaata gcaagaactt ggttgatgta ttcaaaagtg aagaacagcc tcttttgctt
     1081 ttgttgcaaa ttgttttcca acaaaaacat taatttaaca acttctggta cagcaaactg
     1141 gaaacatgca agcacatacc tcacagcaca cgaaaaaagc ccagaacacc tcaattgtat
     1201 gaaagcatgg aaggaactgt cagggaggat cagaagtggg aaaacaattg ataagcagga
     1261 gatggcactt ctggaagagg agcgggtgag atggagagca gtgctaaccc gtctcattgc
     1321 tattgtgcag tcactggcag ttcggaattt ggctctaagg ggacacacag aaacactgtt
     1381 cacatcatca aatgggaatt ttttgaaaga ggttgaactg atggccaggt ttgatcccat
     1441 aatgaaagat catcttaacc gtgtattaag aggaacagca agtcacaaca gctacatagg
     1501 ccatcatgtg cagaatgaac ttattgattt gttgagcagc aaaatcctat ccgctatagt
     1561 ggatgacatc aaaaaggcaa aatatttttc aataattctg gactgcactc tggatataag
     1621 ccacacagaa cagttgtcag ttataattag agtggtgtca ctgatggaga agcctcagat
     1681 cagggaacat tttatggggt ttttggaggc agaggagtcc acaggccagc acttggcatc
     1741 catgatctta aacagacttg aggagttagg aatttctttt gaagactgca gaggacaatc
     1801 atatgataat ggggcaaata tgaaaggcaa aaataaggga gtacaagcca ggctcttaga
     1861 aaagaatccc cgtgctctgt ttttgccatg cggtgcacac acattgaatt tagttgtgtg
     1921 tgatgctgct aagagatctg ttgatgctat gagctacttt ggtgtcctgc aaaagcttta
     1981 cactttattt tcagcctctg cccaacgatg ggccatactg aagagtcagg tgagcatcac
     2041 tctaaagtcg tggacagaaa caaggtggga gagcaaaatc aaaagcatcg agcccatgag
     2101 gtaccaggga gctgcagtga gagaggcttt aatagaagtg agagacaaga ccaaagaccc
     2161 agttataaag gctgaggccc agtctttgtc tgaagaggta gggtcgtacc gcttcaacat
     2221 ctgcacagtc gtatggcatg acattctatc tacaataaag catgtcagca aactcatgca
     2281 gtctccaaat atgcatgtgg acctagctgt gagtcttttg aagaagactg aacaaagtct
     2341 ccagagctac agggcaaatg gctttgtgaa tgcacagatg gcagccaaag aaatgtgcaa
     2401 ggaaatgaat gtcgaggcta ttttgaaaca aaaaagaata agatccacaa agtgccaatt
     2461 ctcgtatgaa tcacacgatg agcctttcag tgacgcactt aaaaagttgg aggttgaatt
     2521 tttcaatgtt gttgttgatg aagccttgtc agccatcgcg gagaggtttt ccacattgga
     2581 agttgtacaa aacagatttg gggttttgac caatttccca agccttggag acgaggagct
     2641 gacggagcaa tgcgaggcac taggcaacat actccatttt gagaagaact gggatttgga
     2701 cagtagagag cttgttcagg aaatcaagaa cttgcctaac ttaccatcaa cgactccaag
     2761 tctccttgag ctcatctctt tcatgtctga taaggatcta tcagaaatct atccgaactt
     2821 ttggactgct ctcaggattg cactcacctt gccagtcact gtggctcaag cagagaggag
     2881 cttttcaaaa ctaaaattga tcaagtcgta cctgaggtca acaatgtcac aggagcgact
     2941 cactaacctt gccgttgtta gcatcaatca ctcagtaggg gagcagatat catatgatga
     3001 tgttattgac gagtttgcat caagaaaggc taggaaggtt aggttttagt tggatccaga
     3061 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3121 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3181 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3241 ggttttttat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa
     3301 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3361 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3421 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3541 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3601 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3901 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3961 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4021 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4081 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4141 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4201 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4261 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4321 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4381 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4441 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4501 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4561 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4621 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4681 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4741 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4801 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4861 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4921 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4981 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5041 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5101 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5161 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5221 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5281 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5461 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5521 tttcgtc
//
