LOCUS       sequence-383804-        4221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway vector containing HA-tagged Creb5.
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(HA)
FEATURES             Location/Qualifiers
     source          1..4221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      311..899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     800..819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1053..1070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1229..1256)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1348..1434)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1483..1505
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1497..1514
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1498..1514
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1530..1629
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             1641..1661
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3201..3227
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3234..3260
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3261..3287
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3307..3406)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3423..3442)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3424..3442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3447..3463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3447..3463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3565..3584)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(3576..4221,1..164)
                     /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(3643..3662)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
       61 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
      121 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
      181 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
      241 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
      301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1081 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     1201 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     1261 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     1321 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     1381 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
     1441 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
     1501 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
     1561 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
     1621 aagcaggctc cgcggccgcc ttgtttaact ttaagaagga gcccttcacc atgatttatg
     1681 aggaatccaa gatgaatttg gagcaggaga ggccgtttgt ctgcagtgcc ccaggctgct
     1741 cccagcgctt cccaacagag gatcatctga tgattcatag gcacaagcat gaaatgactt
     1801 tgaagtttcc ctcaataaaa acagacaata tgttatcaga tcaaactcca accccaacga
     1861 gattcctgaa gaactgcgag gaggtggggc tcttcagtga gttggactgc tccctagagc
     1921 atgagttcag gaaggctcag gaggaagaga gcagcaagcg gaatatctcg atgcataacc
     1981 ccgttggtgg ggccatggcg gggcctggag ctcaccagct cggcagcgcc cggatgccca
     2041 accatgacac cagcgttgtg attcagcaag ccatgccgtc accccagtcc agctctgtca
     2101 tcacgcaggc cccttccacc aaccgccaga tcgggcctgt cccaggctct ctatcttctc
     2161 tgctccatct tcacaacaga cagagacagc ccatgccagc ctccatgcct gggaccctgc
     2221 ccaaccctac aatgccgggc tcttctgccg tcttgatgcc aatggagaga caaatgtcag
     2281 tgaactccaa cctcctggga atgcaaggtc caaatctcag caacccctgt gcttctcccc
     2341 aagtccagcc aatgcattca gaagctaaga tgaggctgaa ggctgcattg actcaccacc
     2401 ctgctgccat gtcgaatggg aatatgaaca ccatgggcca catgatggaa atgatgggct
     2461 cccggcagga ccagacgcca caccatcaca tgcactcaca cccgcatcag caccagacac
     2521 tgccggccca ccacccctac ccgcaccagc accagcaccc agcacaccac cctcatcctc
     2581 aaccccatca ccagcagaac cacccacatc atcactccca ctcccacctt catgcacacc
     2641 cagcacatca ccagacctcg ccacacccac ccctgcactc cggcacccaa gcacaggttt
     2701 caccagcaac acaacagatg cagcccaccc agacaataca gccgccccag cccacagggg
     2761 ggcgccggcg aagggtggtg gatgaggatc ctgacgagag gcggcggaaa tttctggaac
     2821 ggaaccgggc ggccgccacc cgctgcaggc agaagaggaa ggtctgggtg atgtcactgg
     2881 aaaagaaagc agaagagctc acccagacaa acatgcagct tcagaacgaa gtgtccatgt
     2941 tgaagaacga ggtggcacag ctgaagcagc tgttgttaac acataaagac tgccccataa
     3001 cagccatgca gaaagaatca caaggatatc tcagtccaga gagtagccct cctgcaagtc
     3061 ccacccccgc atgctcacag cagcaggtca tccagcataa caccatcacc acttcctcgg
     3121 cagtcagcga ggtggtagga agctctaccc tcagccagct caccactcac agaacagacc
     3181 tgaaccccat cctcggaggt tacccttacg acgtcccaga ctacgctggc tcctaccctt
     3241 acgacgtccc agactacgct tacccttacg acgtcccaga ctacgcttaa aagggtgggc
     3301 gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     3361 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     3421 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     3481 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     3541 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     3601 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     3661 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     3721 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     3781 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     3841 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     3901 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     3961 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     4021 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     4081 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     4141 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     4201 taataggttg tattgatgtt g
//
