LOCUS       sequence-379111-        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  JOURNAL   Unpublished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            "Unpublished"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_NN_G1333-DddA11-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     369..386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(545..572)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(664..750)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     814..830
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    847..942
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             945..989
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1299..1709)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1574..1590
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1600..1618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1666..1686)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1666..1684)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1705..1721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1718..1740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1729..1745
                     /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(1753..1759,1760..1777,1778..1783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1798..1819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2191..2442
                     /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(2464..2563)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2594..2612)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2617..2633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2735..2754)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2746..3555
                     /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(2813..2832)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3702..4075,1..215)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
       61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      481 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      541 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      601 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      661 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      721 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      781 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      841 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      901 agcaatgctt ttttataatg ccaactttgt ataataaagt tgataaggag accgccgctg
      961 ccaagttcga gagacagcac atggacagca tcgatatcgc cgatctacgc acgctcggct
     1021 acagccagca gcaacaggag aagatcaaac cgaaggttcg ttcgacagtg gcgcagcacc
     1081 acgaggcact ggtcggccac gggtttacac acgcgcacat cgttgcgtta agccaacacc
     1141 cggcagcgtt agggaccgtc gctgtcaagt atcaggacat gatcgcagcg ttgccagagg
     1201 cgacacacga agcgatcgtt ggcgtcggca aacagtggtc cggcgcacgc gctctggagg
     1261 ccttgctcac ggtggcggga gagttgagag gtccaccgtt acagttggac acaggccaac
     1321 ttctcaagat tgcaaaacgt ggcggcgtga ccgcagtgga ggcagtgcat gcatggcgca
     1381 atgcactgac gggtgccccg ctcaaggaga cgggcgccgc tacagggcgc gtcccattcg
     1441 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     1501 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     1561 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     1621 gaattgggta ccgggccccc cctcgaggtc ctccagcttt tgttcccttt agtgagggtt
     1681 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1741 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1801 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccacc ggtcgtctcc
     1861 acccctgagc aggtagtggc tattgcatcc aacaacgggg gcagacccgc actggagtca
     1921 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
     1981 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
     2041 ggatctggta gctatgcgtt aggaccctat cagatttcag ctcctcaatt gcctgcctat
     2101 aatgggcaaa ctgttggcac cttttactac gtcaatgatg ctggagggtt agaatccaag
     2161 gtgttcataa gtggtggttc tggaggtagt acgaatcttt cggacatcat agagaaggaa
     2221 actggaaaac agctcgttat ccaagagagc attctcatgt tgccagaaga agttgaagag
     2281 gttataggca acaaaccgga atctgacatt ctggtacata ccgcttatga tgagtcaaca
     2341 gatgagaacg tcatgctttt gacatctgat gcaccagaat acaaaccttg ggcacttgtg
     2401 attcaggatt ccaatggtga gaacaagatc aagatgctat gataactcga gcgatcctct
     2461 agaacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2521 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagggc
     2581 ggccgcgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     2641 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2701 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2761 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2821 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     2881 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2941 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     3001 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     3061 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     3121 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     3181 cgccttgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     3241 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     3301 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3361 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     3421 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3481 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     3541 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     3601 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     3661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3841 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actga
//
