LOCUS       sequence-379112-        4075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD_v2mod vector assembly, Entry clone 1 with attL2 and L3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4075)
  AUTHORS   Shin-ichi Arimura Lab
  TITLE     Shin-ichi Arimura Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4075)
  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: 
            "Unpublished"
COMMENT     Sequence Label: E3_pENTR_L3-L2_NI_G1333-DddA11-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    864..959
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             962..1006
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1316..1726)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1591..1607
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1617..1635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1683..1703)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1683..1701)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1722..1738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1735..1757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1746..1762
                     /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(1770..1776,1777..1794,1795..1800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1815..1836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2208..2459
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    complement(2481..2580)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2611..2629)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2634..2650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2752..2771)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2763..3572
                     /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     complement(2830..2849)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3719..4075,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtata ataaagttga
      961 taaggagacc gccgctgcca agttcgagag acagcacatg gacagcatcg atatcgccga
     1021 tctacgcacg ctcggctaca gccagcagca acaggagaag atcaaaccga aggttcgttc
     1081 gacagtggcg cagcaccacg aggcactggt cggccacggg tttacacacg cgcacatcgt
     1141 tgcgttaagc caacacccgg cagcgttagg gaccgtcgct gtcaagtatc aggacatgat
     1201 cgcagcgttg ccagaggcga cacacgaagc gatcgttggc gtcggcaaac agtggtccgg
     1261 cgcacgcgct ctggaggcct tgctcacggt ggcgggagag ttgagaggtc caccgttaca
     1321 gttggacaca ggccaacttc tcaagattgc aaaacgtggc ggcgtgaccg cagtggaggc
     1381 agtgcatgca tggcgcaatg cactgacggg tgccccgctc aaggagacgg gcgccgctac
     1441 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1501 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1561 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1621 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcctc cagcttttgt
     1681 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     1741 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1801 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1861 ttccaccggt cgtctccacc cctgagcagg tagtggctat tgcatccaac atagggggca
     1921 gacccgcact ggagtcaatt gttgcccagt tatctcgccc tgatccggcg ttggccgcgt
     1981 tgaccaacga ccacctcgtc gccttggcct gcctcggcgg gcgtcctgcg ctggatgcag
     2041 tgaaaaaggg attgggggga tctggtagct atgcgttagg accctatcag atttcagctc
     2101 ctcaattgcc tgcctataat gggcaaactg ttggcacctt ttactacgtc aatgatgctg
     2161 gagggttaga atccaaggtg ttcataagtg gtggttctgg aggtagtacg aatctttcgg
     2221 acatcataga gaaggaaact ggaaaacagc tcgttatcca agagagcatt ctcatgttgc
     2281 cagaagaagt tgaagaggtt ataggcaaca aaccggaatc tgacattctg gtacataccg
     2341 cttatgatga gtcaacagat gagaacgtca tgcttttgac atctgatgca ccagaataca
     2401 aaccttgggc acttgtgatt caggattcca atggtgagaa caagatcaag atgctatgat
     2461 aactcgagcg atcctctaga acccagcttt cttgtacaaa gttggcatta taagaaagca
     2521 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2581 ccatccagct gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     2641 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2701 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2761 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2821 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2881 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2941 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3001 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3061 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3121 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3181 acagcgatcg cgtatttcgc cttgctcagg cgcaatcacg aatgaataac ggtttggttg
     3241 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3301 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3361 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3421 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3481 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3541 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3601 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3661 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3721 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3781 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3841 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3901 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3961 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4021 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaac
//
