LOCUS       sequence-244265-        4145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full MCS (step 2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4145)
  AUTHORS   Santos-Moreno J, Schaerli Y
  TITLE     A Framework for the Modular and Combinatorial Assembly of Synthetic 
            Gene Circuits.
  JOURNAL   ACS Synth Biol. 2019 Jul 19;8(7):1691-1697. doi: 
            10.1021/acssynbio.9b00174. Epub 2019 Jun 24.
  PUBMED    31185158
REFERENCE   2  (bases 1 to 4145)
  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"; doi: 
            "10.1021/acssynbio.9b00174"; journalName: "ACS Synth Biol"; date: 
            "2019-07-19- 19"; volume: "8"; issue: "7"; pages: "1691-1697"
COMMENT     Sequence Label: pC-0_v2
FEATURES             Location/Qualifiers
     source          1..4145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..311
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     234..253
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      1346..1373
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(1498..2289,2290..2358))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2121..2140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2359..2463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2481..3116)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(2626..2645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3267..4145)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
       61 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
      121 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
      181 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
      241 agcattttta tccataagat tagcggttcc tacctgacgc tttttatcgc aactctctac
      301 tgtttctcca taccgaattc ataggataga ttctggaaac tttaccgtcc gagctcaggc
      361 ttaccttact cgagcaataa acagttgata gggcttctcc gttaccatgg ttcagccaaa
      421 aaacttaaga ccgccggtct tgtccactac cttgcagtaa tgcggtggac aggatcggcg
      481 gttttctttt ctcttctcaa ttcttctgac ctgtaacgaa taatagatag taaagtagtc
      541 tccgattgag ttttctctgc cgagtcccac ccagttctgt gatttcagta agttggtaat
      601 tgatacactg ttgcgagaac tgctgcctgg tagtagatag gttgttattg agtaagaagg
      661 taaagtgaac gaaatccctg aaactgagac tgtagaaaat aagcttgtcc agactattgg
      721 atccaagaga tttctacacg attgagcact gtctcctgca ggctcggtac caaattccag
      781 aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg tcctaataga taaaggatag
      841 gtctggtagt gttgttcgtt ctcgcaggta aatcaataat actcagcagt tccgtagact
      901 tttcagtggg acagggtagc gataacagat agattgtaat aagacacagt aggtgctcgt
      961 agttgcgtga agagaaccgc tcaggaaatc cagtcagaag tattggtaat cgttgaaaac
     1021 tcagtcgacg cacttactga agacgtccta ttacactcgt cgttggaaac tgaagatgcg
     1081 gccgcggaaa cacagaaaaa agcccgcacc tgacagtgcg ggcttttttt ttcgaccaaa
     1141 ggtagcgaac gacgagtcac tgttgaggat aaatactttc tctactaggc gcctgttaca
     1201 caggtcctca gcggcgcgcc tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1261 ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     1321 taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt
     1381 cagatctggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1441 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1501 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1561 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1621 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1681 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1741 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1801 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1861 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1921 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1981 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2041 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2101 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2161 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2221 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2281 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2341 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2461 gcgcagcatg ctagcggcag aaacgtccta gaagatgcca ggaggatact tagcagagag
     2521 acaataaggc cggagcgaag ccgtttttcc ataggctccg cccccctgac gaacatcacg
     2581 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     2641 ttccccctga tggctccctc ttgcgctctc ctgttcccgt cctgcggcgt ccgtgttgtg
     2701 gtggaggctt tacccaaatc accacgtccc gttccgtgta gacagttcgc tccaagctgg
     2761 gctgtgtgca agaacccccc gttcagcccg actgctgcgc cttatccggt aactatcatc
     2821 ttgagtccaa cccggaaaga cacgacaaaa cgccactggc agcagccatt ggtaactgag
     2881 aattagtgga tttagatatc gagagtcttg aagtggtggc ctaacagagg ctacactgaa
     2941 aggacagtat ttggtatctg cgctccacta aagccagtta ccaggttaag cagttcccca
     3001 actgacttaa ccttcgatca aaccgcctcc ccaggcggtt ttttcgttta cagagcagga
     3061 gattacgacg atcgtaaaag gatctcaaga agatccttta cggattcccg acaccatcac
     3121 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     3181 ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     3241 gctcctaggg tctgattcgt taccaattat gacaacttga cggctacatc attcactttt
     3301 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     3361 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     3421 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     3481 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     3541 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     3601 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     3661 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     3721 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     3781 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     3841 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     3901 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     3961 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     4021 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     4081 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     4141 gccat
//
