LOCUS       sequence-178094-        4146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            TCF7 CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4146)
  AUTHORS   Jackel C, Nogueira MS, Ehni N, Kraus C, Ranke J, Dohmann M, Noessner
            E, Nelson PJ
  TITLE     A vector platform for the rapid and efficient engineering of stable 
            complex transgenes.
  JOURNAL   Sci Rep. 2016 Oct 3;6:34365. doi: 10.1038/srep34365.
  PUBMED    27694838
REFERENCE   2  (bases 1 to 4146)
  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: "Sci Rep."; 
            date: "2016-10-3"; volume: "6:34365. doi"; pages: " 
            10.1038/srep34365"
COMMENT     Sequence Label: pENTR221-TCF7
FEATURES             Location/Qualifiers
     source          1..4146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             728..754
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1910..1943
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(1978..1996)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1979..1996)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2016..2033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2022..2246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(2258..2357)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2387..2406)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2388..2406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2411..2427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2411..2427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2529..2548)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2540..3349
                     /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(2607..2626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3985..4004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttgtaagc tcggatccac tagtaacggc cgccagtgtg ctggaattct gcagatcggc
      721 caccatgtat ccatatgacg tcccagacta tgccatgccg cagctggact ccggcggggg
      781 cggcgcgggc ggcggcgacg acctcggcgc gccggacgag ctgctggcct tccaggatga
      841 aggcgaggag caggacgaca agagccgcga cagcgccgcc ggtcccgagc gcgacctggc
      901 cgagctcaag tcgtcgctcg tgaacgagtc cgagggcgcg gcaggcggcg cagggatccc
      961 gggggtcccg ggggccggcg ccggggcccg cggcgaggcc gaggctctcg ggcgggaaca
     1021 cgctgcgcag agactcttcc cggacaaact tccagagccc ctggaggacg gcctgaaggc
     1081 cccggagtgc accagcggca tgtacaaaga gaccgtctac tccgccttca atctgctcat
     1141 gcattaccca cccccctcgg gagcagggca gcacccccag ccgcagcccc cgctgcacaa
     1201 ggccaatcag cccccccacg gtgtccccca actctctctc tacgaacatt tcaacagccc
     1261 acatcccacc cctgcacctg cggacatcag ccagaagcaa gttcacaggc ctctgcagac
     1321 ccctgacctc tctggcttct actccctgac ctcaggcagc atggggcagc tcccccacac
     1381 tgtgagctgg ttcacccacc catccttgat gctaggttct ggtgtacctg gtcacccagc
     1441 agccatcccc cacccggcca ttgtgccccc ctcagggaag caggagctgc agcccttcga
     1501 ccgcaacctg aagacacaag cagagtccaa ggcagagaag gaggccaaga agccaaccat
     1561 caagaagccc ctcaatgcct tcatgctgta catgaaggag atgagagcca aggtcattgc
     1621 agagtgcaca cttaaggaga gcgctgccat caaccagatc ctgggccgca ggtggcacgc
     1681 gctgtcgcga gaagagcagg ccaagtacta tgagctggcc cgcaaggaga ggcagctgca
     1741 catgcagcta tacccaggct ggtcagcgcg ggacaactac gggaagaaga agaggcggtc
     1801 gagggaaaag caccaagaat ccaccacagg aggaaaaaga aatgcattcg gtacttaccc
     1861 ggagaaggcc gctgccccag ccccgttcct tccgatgaca gtgctctaga taacttcgta
     1921 taatgtatgc tatacgaagt tatgcggccg ctcgagcatg catctagagg gccctattct
     1981 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     2041 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2101 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2161 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2221 tgctggggat gcggtgggct ctatggcttc tgaggcaacc cagctttctt gtacaaagtt
     2281 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2341 aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct atagtgagtc
     2401 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2461 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2521 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2581 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2641 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2701 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2761 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2821 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2881 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2941 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3001 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3061 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3121 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3181 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3241 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3301 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3361 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3421 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgtt
//
