LOCUS       sequence-383807-        4221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway vector containing HA-tagged Creb5 with T59/T61D 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/T61D)(HA)
FEATURES             Location/Qualifiers
     source          1..4221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     241..260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     494..511
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(670..697)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(789..875)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     924..946
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     938..955
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     939..955
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    971..1070
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             1082..1102
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2642..2668
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2675..2701
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2702..2728
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2748..2847)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2864..2883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2865..2883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2888..2904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2888..2904)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3006..3025)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3017..3826
                     /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(3084..3103)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3973..4221,1..340)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
       61 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      121 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      181 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      241 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      301 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      361 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      421 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
      481 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
      541 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      601 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
      661 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
      721 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
      781 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
      841 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
      901 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
      961 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
     1021 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc
     1081 cttgtttaac tttaagaagg agcccttcac catgatttat gaggaatcca agatgaattt
     1141 ggagcaggag aggccgtttg tctgcagtgc cccaggctgc tcccagcgct tcccaacaga
     1201 ggatcatctg atgattcata ggcacaagca tgaaatgact ttgaagtttc cctcaataaa
     1261 aacagacaat atgttatcag atcaagatcc agacccaacg agattcctga agaactgcga
     1321 ggaggtgggg ctcttcagtg agttggactg ctccctagag catgagttca ggaaggctca
     1381 ggaggaagag agcagcaagc ggaatatctc gatgcataac cccgttggtg gggccatggc
     1441 ggggcctgga gctcaccagc tcggcagcgc ccggatgccc aaccatgaca ccagcgttgt
     1501 gattcagcaa gccatgccgt caccccagtc cagctctgtc atcacgcagg ccccttccac
     1561 caaccgccag atcgggcctg tcccaggctc tctatcttct ctgctccatc ttcacaacag
     1621 acagagacag cccatgccag cctccatgcc tgggaccctg cccaacccta caatgccggg
     1681 ctcttctgcc gtcttgatgc caatggagag acaaatgtca gtgaactcca acctcctggg
     1741 aatgcaaggt ccaaatctca gcaacccctg tgcttctccc caagtccagc caatgcattc
     1801 agaagctaag atgaggctga aggctgcatt gactcaccac cctgctgcca tgtcgaatgg
     1861 gaatatgaac accatgggcc acatgatgga aatgatgggc tcccggcagg accagacgcc
     1921 acaccatcac atgcactcac acccgcatca gcaccagaca ctgccggccc accaccccta
     1981 cccgcaccag caccagcacc cagcacacca ccctcatcct caaccccatc accagcagaa
     2041 ccacccacat catcactccc actcccacct tcatgcacac ccagcacatc accagacctc
     2101 gccacaccca cccctgcact ccggcaccca agcacaggtt tcaccagcaa cacaacagat
     2161 gcagcccacc cagacaatac agccgcccca gcccacaggg gggcgccggc gaagggtggt
     2221 ggatgaggat cctgacgaga ggcggcggaa atttctggaa cggaaccggg cggccgccac
     2281 ccgctgcagg cagaagagga aggtctgggt gatgtcactg gaaaagaaag cagaagagct
     2341 cacccagaca aacatgcagc ttcagaacga agtgtccatg ttgaagaacg aggtggcaca
     2401 gctgaagcag ctgttgttaa cacataaaga ctgccccata acagccatgc agaaagaatc
     2461 acaaggatat ctcagtccag agagtagccc tcctgcaagt cccacccccg catgctcaca
     2521 gcagcaggtc atccagcata acaccatcac cacttcctcg gcagtcagcg aggtggtagg
     2581 aagctctacc ctcagccagc tcaccactca cagaacagac ctgaacccca tcctcggagg
     2641 ttacccttac gacgtcccag actacgctgg ctcctaccct tacgacgtcc cagactacgc
     2701 ttacccttac gacgtcccag actacgctta aaagggtggg cgcgccgacc cagctttctt
     2761 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2821 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2881 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2941 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3001 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     3061 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     3121 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3181 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3241 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3301 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3361 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3421 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3481 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3541 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3601 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3661 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3721 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3781 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3841 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3901 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3961 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4021 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4081 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4141 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4201 accagtggct gctgccagtg g
//
