LOCUS       sequence-379205-        4202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD_v2mod vector assembly, Entry clone 1 with attL1 and L4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4202)
  AUTHORS   Shin-ichi Arimura Lab
  TITLE     Shin-ichi Arimura Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4202)
  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            "Unpublished"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: E1_pENTR_L1-L4_NG_G1333-DddA11-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(541..568)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(660..746)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     810..826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    842..941
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             944..970
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1277..1687)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1552..1568
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1578..1596
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1644..1664)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1644..1662)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1683..1699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1696..1718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1707..1723
                     /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(1731..1737,1738..1755,1756..1761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1776..1797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2319..2570
                     /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(2596..2691)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2717..2735)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2740..2756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2858..2877)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2869..3678
                     /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(2936..2955)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3825..4202,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcatggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
      541 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
      601 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
      661 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      721 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      781 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      841 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
      901 atgctttttt ataatgccaa ctttgtacaa aaaagcaggc ttttacccat acgatgttcc
      961 agattacgct gatatcgccg atctacgcac gctcggctac agccagcagc aacaggagaa
     1021 gatcaaaccg aaggttcgtt cgacagtggc gcagcaccac gaggcactgg tcggccacgg
     1081 gtttacacac gcgcacatcg ttgcgttaag ccaacacccg gcagcgttag ggaccgtcgc
     1141 tgtcaagtat caggacatga tcgcagcgtt gccagaggcg acacacgaag cgatcgttgg
     1201 cgtcggcaaa cagtggtccg gcgcacgcgc tctggaggcc ttgctcacgg tggcgggaga
     1261 gttgagaggt ccaccgttac agttggacac aggccaactt ctcaagattg caaaacgtgg
     1321 cggcgtgacc gcagtggagg cagtgcatgc atggcgcaat gcactgacgg gtgccccgct
     1381 caaggagacg ggcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     1441 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     1501 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     1561 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     1621 tcgaggtcct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     1681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1801 gcgttgcgct cactgcccgc tttccaccgg tcgtctccac ccctgagcag gtagtggcta
     1861 ttgcatccaa cggagggggc agacccgcac tggagtcaat tgttgcccag ttatctcgcc
     1921 ctgatccggc gttggccgcg ttgaccaacg accacctcgt cgccttggcc tgcctcggcg
     1981 ggcgtcctgc gctggatgca gtgaaaaagg gattgggggg tagtccaact ccatatccca
     2041 attatgtgag tgcaggccat gttgaaggtc aatcagccct attcatgaga gataacggaa
     2101 taagtgaagg actggtgttt cacaacaatc caaaaggtac ttgtggattt tgcgtaaaca
     2161 tgattgaaac ccttctccca gaaaacgcta agatgacagt agttccacct gaaggtgcta
     2221 ttcctgtcaa aagaggtgct actggtgaaa ccaaggtgtt tattgggaat tccaattcac
     2281 ccaaaagccc aacgaaaggt gggtgtagtg gaggatctac aaatctctct gacatcattg
     2341 agaaagagac tggaaagcaa ctagtcattc aggagtcaat cctgatgtta ccagaggagg
     2401 ttgaggaagt gataggcaat aagcccgaaa gcgatatact tgttcatact gcctatgacg
     2461 aatcgacgga tgagaacgta atgcttctaa cctcagatgc tcctgagtac aaaccttggg
     2521 cgttagttat ccaggattcc aatggagaga acaagatcaa gatgttgtga taactcgagc
     2581 gatcctctag acacccaact tttctataca aagttggcat tataagaaag cattgcttat
     2641 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     2701 ctgcagggcg gccgccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     2761 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     2821 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     2881 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     2941 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     3001 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     3061 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     3121 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     3181 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     3241 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     3301 tttcgccttg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     3361 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     3421 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     3481 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     3541 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     3601 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     3661 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     3721 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     3781 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3841 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3901 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3961 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4021 ccgcctacat accttgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4081 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4141 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4201 ta
//
