LOCUS       sequence-383806-        4221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway vector containing HA-tagged Creb5 with T59/T61A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4221)
  AUTHORS   Zhang CH, Gao Y, Jadhav U, Hung HH, Holton KM, Grodzinsky AJ, 
            Shivdasani RA, Lassar AB
  TITLE     Creb5 establishes the competence for Prg4 expression in articular 
            cartilage.
  JOURNAL   Commun Biol. 2021 Mar 12;4(1):332. doi: 10.1038/s42003-021-01857-0.
  PUBMED    33712729
REFERENCE   2  (bases 1 to 4221)
  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: "Commun 
            Biol."; date: "2021-03-12"; volume: "4(1):332. doi"; pages: " 
            10.1038/s42003-021-01857-0"
COMMENT     Sequence Label: pENTR-Creb5(T59/T61A)(HA)
FEATURES             Location/Qualifiers
     source          1..4221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(147..174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(266..352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     401..423
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     415..432
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     416..432
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    448..547
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             559..579
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2119..2145
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2152..2178
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2179..2205
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2225..2324)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2341..2360)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2342..2360)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2365..2381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2365..2381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2483..2502)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2494..3303
                     /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(2561..2580)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3450..4038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3939..3958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4192..4209
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      121 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
      241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
      301 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
      361 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
      421 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
      481 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      541 gcaggctccg cggccgcctt gtttaacttt aagaaggagc ccttcaccat gatttatgag
      601 gaatccaaga tgaatttgga gcaggagagg ccgtttgtct gcagtgcccc aggctgctcc
      661 cagcgcttcc caacagagga tcatctgatg attcataggc acaagcatga aatgactttg
      721 aagtttccct caataaaaac agacaatatg ttatcagatc aagctccagc cccaacgaga
      781 ttcctgaaga actgcgagga ggtggggctc ttcagtgagt tggactgctc cctagagcat
      841 gagttcagga aggctcagga ggaagagagc agcaagcgga atatctcgat gcataacccc
      901 gttggtgggg ccatggcggg gcctggagct caccagctcg gcagcgcccg gatgcccaac
      961 catgacacca gcgttgtgat tcagcaagcc atgccgtcac cccagtccag ctctgtcatc
     1021 acgcaggccc cttccaccaa ccgccagatc gggcctgtcc caggctctct atcttctctg
     1081 ctccatcttc acaacagaca gagacagccc atgccagcct ccatgcctgg gaccctgccc
     1141 aaccctacaa tgccgggctc ttctgccgtc ttgatgccaa tggagagaca aatgtcagtg
     1201 aactccaacc tcctgggaat gcaaggtcca aatctcagca acccctgtgc ttctccccaa
     1261 gtccagccaa tgcattcaga agctaagatg aggctgaagg ctgcattgac tcaccaccct
     1321 gctgccatgt cgaatgggaa tatgaacacc atgggccaca tgatggaaat gatgggctcc
     1381 cggcaggacc agacgccaca ccatcacatg cactcacacc cgcatcagca ccagacactg
     1441 ccggcccacc acccctaccc gcaccagcac cagcacccag cacaccaccc tcatcctcaa
     1501 ccccatcacc agcagaacca cccacatcat cactcccact cccaccttca tgcacaccca
     1561 gcacatcacc agacctcgcc acacccaccc ctgcactccg gcacccaagc acaggtttca
     1621 ccagcaacac aacagatgca gcccacccag acaatacagc cgccccagcc cacagggggg
     1681 cgccggcgaa gggtggtgga tgaggatcct gacgagaggc ggcggaaatt tctggaacgg
     1741 aaccgggcgg ccgccacccg ctgcaggcag aagaggaagg tctgggtgat gtcactggaa
     1801 aagaaagcag aagagctcac ccagacaaac atgcagcttc agaacgaagt gtccatgttg
     1861 aagaacgagg tggcacagct gaagcagctg ttgttaacac ataaagactg ccccataaca
     1921 gccatgcaga aagaatcaca aggatatctc agtccagaga gtagccctcc tgcaagtccc
     1981 acccccgcat gctcacagca gcaggtcatc cagcataaca ccatcaccac ttcctcggca
     2041 gtcagcgagg tggtaggaag ctctaccctc agccagctca ccactcacag aacagacctg
     2101 aaccccatcc tcggaggtta cccttacgac gtcccagact acgctggctc ctacccttac
     2161 gacgtcccag actacgctta cccttacgac gtcccagact acgcttaaaa gggtgggcgc
     2221 gccgacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2281 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2341 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2401 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2461 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2521 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2581 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2641 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2701 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2761 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2821 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2881 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2941 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     3001 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     3061 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     3121 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3181 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3241 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3301 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3361 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3421 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3481 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3541 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3601 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3661 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3721 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3781 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3841 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3901 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3961 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4021 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4081 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4141 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4201 gtgagcgagg aagcggaaga g
//
