LOCUS       sequence-379113-        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  JOURNAL   Unpublished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            "Unpublished"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_NG_G1333-DddA11-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..126
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(436..846)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     711..727
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        737..755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(803..823)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(803..821)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(842..858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(855..877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    866..882
                     /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(890..896,897..914,915..920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    935..956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1328..1579
                     /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(1601..1700)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1731..1749)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1754..1770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1872..1891)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1883..2692
                     /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(1950..1969)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2839..3427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3328..3347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3581..3598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3757..3784)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3876..3962)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4026..4042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(4059..4075,1..79)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
       61 actttgtata ataaagttga taaggagacc gccgctgcca agttcgagag acagcacatg
      121 gacagcatcg atatcgccga tctacgcacg ctcggctaca gccagcagca acaggagaag
      181 atcaaaccga aggttcgttc gacagtggcg cagcaccacg aggcactggt cggccacggg
      241 tttacacacg cgcacatcgt tgcgttaagc caacacccgg cagcgttagg gaccgtcgct
      301 gtcaagtatc aggacatgat cgcagcgttg ccagaggcga cacacgaagc gatcgttggc
      361 gtcggcaaac agtggtccgg cgcacgcgct ctggaggcct tgctcacggt ggcgggagag
      421 ttgagaggtc caccgttaca gttggacaca ggccaacttc tcaagattgc aaaacgtggc
      481 ggcgtgaccg cagtggaggc agtgcatgca tggcgcaatg cactgacggg tgccccgctc
      541 aaggagacgg gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
      601 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      661 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      721 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
      781 cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
      841 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      901 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      961 cgttgcgctc actgcccgct ttccaccggt cgtctccacc cctgagcagg tagtggctat
     1021 tgcatccaac ggagggggca gacccgcact ggagtcaatt gttgcccagt tatctcgccc
     1081 tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct gcctcggcgg
     1141 gcgtcctgcg ctggatgcag tgaaaaaggg attgggggga tctggtagct atgcgttagg
     1201 accctatcag atttcagctc ctcaattgcc tgcctataat gggcaaactg ttggcacctt
     1261 ttactacgtc aatgatgctg gagggttaga atccaaggtg ttcataagtg gtggttctgg
     1321 aggtagtacg aatctttcgg acatcataga gaaggaaact ggaaaacagc tcgttatcca
     1381 agagagcatt ctcatgttgc cagaagaagt tgaagaggtt ataggcaaca aaccggaatc
     1441 tgacattctg gtacataccg cttatgatga gtcaacagat gagaacgtca tgcttttgac
     1501 atctgatgca ccagaataca aaccttgggc acttgtgatt caggattcca atggtgagaa
     1561 caagatcaag atgctatgat aactcgagcg atcctctaga acccagcttt cttgtacaaa
     1621 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1681 caaaataaaa tcattatttg ccatccagct gcagggcggc cgcgatatcc cctatagtga
     1741 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1801 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1861 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1921 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1981 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2041 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2101 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2161 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2221 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2281 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc cttgctcagg cgcaatcacg
     2341 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2401 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2461 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2521 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2581 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2641 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2701 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2761 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3541 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3601 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3661 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     3721 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3781 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     3841 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     3901 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     3961 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
     4021 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattt
//
