LOCUS       sequence-178935-        3149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning of a COL2A1-based, chondrogenesis-responsive promoter, 
            and subcloning this insert into new vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3149)
  AUTHORS   De La Vega RE, Scheu M, Brown LA, Evans C, Ferreira E, Porter RM
  TITLE     Specific, sensitive and stable reporting of human mesenchymal 
            stromal cell chondrogenesis.
  JOURNAL   Tissue Eng Part C Methods. 2019 Feb 6. doi: 
            10.1089/ten.TEC.2018.0295.
  PUBMED    30727864
REFERENCE   2  (bases 1 to 3149)
  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: "Tissue Eng 
            Part C Methods. 2019 Feb 6. doi: 10.1089/ten.TEC.2018.0295."
COMMENT     Sequence Label: pUC57-4eCOL2A1
FEATURES             Location/Qualifiers
     source          1..3149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(941..963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    952..968
                     /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(976..982,983..1000,1001..1006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1021..1042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1159..1176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1330..1918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1410..1429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2089..2880,2881..2949))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2712..2731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2950..3054)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3122..3140
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tgtgtcccaa tggatccgaa gcgggagacc cggcagccca gcagcgctct
      481 gcgtcttctc cccgccgcgg cgcccgttat atgcgcccgg cccctttcga ggctggcgaa
      541 ctcgcccaaa ccgcgggccg ccccctgccc ccagctgggc tgtaacctga accgcccgcc
      601 cccggagccc gcctgggccc tgccagtgcc cgcacctgcc caagccggct agctctcatg
      661 aatggggctt ttctcgagcg cacacagagc ccgattcaca gctctcatga atggggcttt
      721 tctcgagcgc acacagagcc cgattcacag ctctcatgaa tggggctttt ctcgagcgca
      781 cacagagccc gattcacagc tctcatgaat ggggcttttc tcgagcgcac acagagcccg
      841 attcacagac tagtagatct gtgtcattgg atcggatccc gggcccgtcg actgcagagg
      901 cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1741 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2221 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2941 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3001 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3061 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3121 aaataggcgt atcacgaggc cctttcgtc
//
