LOCUS       sequence-244266-        4150 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 4150)
  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 4150)
  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
FEATURES             Location/Qualifiers
     source          1..4150
                     /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(1561..2376)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1561..2376)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2284..2303
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2377..2468)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2486..3121)
                     /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(2631..2650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3272..4150)
                     /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 cagatctacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
     1441 cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga
     1501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     1561 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     1621 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     1681 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     1741 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     1801 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     1861 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     1921 cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa caggaatcga
     1981 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     2041 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     2101 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     2161 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     2221 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     2281 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     2341 cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg
     2401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2461 taaacaaata ggcatgctag cgcagaaacg tcctagaaga tgccaggagg atacttagca
     2521 gagagacaat aaggccggag cgaagccgtt tttccatagg ctccgccccc ctgacgaaca
     2581 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     2641 ggcgtttccc cctgatggct ccctcttgcg ctctcctgtt cccgtcctgc ggcgtccgtg
     2701 ttgtggtgga ggctttaccc aaatcaccac gtcccgttcc gtgtagacag ttcgctccaa
     2761 gctgggctgt gtgcaagaac cccccgttca gcccgactgc tgcgccttat ccggtaacta
     2821 tcatcttgag tccaacccgg aaagacacga caaaacgcca ctggcagcag ccattggtaa
     2881 ctgagaatta gtggatttag atatcgagag tcttgaagtg gtggcctaac agaggctaca
     2941 ctgaaaggac agtatttggt atctgcgctc cactaaagcc agttaccagg ttaagcagtt
     3001 ccccaactga cttaaccttc gatcaaaccg cctccccagg cggttttttc gtttacagag
     3061 caggagatta cgacgatcgt aaaaggatct caagaagatc ctttacggat tcccgacacc
     3121 atcactctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     3181 acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact
     3241 gggttgctcc tagggtctga ttcgttacca attatgacaa cttgacggct acatcattca
     3301 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     3361 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     3421 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     3481 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     3541 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     3601 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     3661 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     3721 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     3781 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     3841 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     3901 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     3961 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     4021 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     4081 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     4141 cttcagccat
//
