LOCUS       sequence-181460-        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(531..548)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(531..547)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(540..562)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(756..775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     875..897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(935..953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1021..1125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1126..1194,1195..1986)
                     /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     complement(1344..1363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2157..2745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2646..2665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2899..2916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3033..3054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3069..3074,3075..3092,3093..3099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3107..3123
                     /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)."
     primer_bind     3112..3134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3131..3147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3131..3147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgattacgcc aagcttgcat gcaggcctct gcagtcgacg ggcccgggat ccgatccaat
       61 gacacagatc tactagtctg tgaatcgggc tctgtgtgcg ctcgagaaaa gccccattca
      121 tgagagctgt gaatcgggct ctgtgtgcgc tcgagaaaag ccccattcat gagagctgtg
      181 aatcgggctc tgtgtgcgct cgagaaaagc cccattcatg agagctgtga atcgggctct
      241 gtgtgcgctc gagaaaagcc ccattcatga gagctagccg gcttgggcag gtgcgggcac
      301 tggcagggcc caggcgggct ccgggggcgg gcggttcagg ttacagccca gctgggggca
      361 gggggcggcc cgcggtttgg gcgagttcgc cagcctcgaa aggggccggg cgcatataac
      421 gggcgccgcg gcggggagaa gacgcagagc gctgctgggc tgccgggtct cccgcttcgg
      481 atccattggg acacatctag atgcattcgc gaggtaccga gctcgaattc actggccgtc
      541 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      601 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      661 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
      721 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
      781 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
      841 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
      901 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
      961 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1021 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1081 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1141 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1201 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1261 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1321 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1381 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1441 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1501 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1561 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1621 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1681 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1741 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1801 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1861 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1921 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1981 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2041 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2161 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2221 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2281 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2341 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2401 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2461 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2521 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2581 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2641 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2701 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2761 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2821 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2881 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2941 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3001 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     3061 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     3121 caatttcaca caggaaacag ctatgacca
//
