LOCUS       sequence-244243-        5403 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 5403)
  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 5403)
  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_v3
FEATURES             Location/Qualifiers
     source          1..5403
                     /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"
     primer_bind     complement(1229..1247)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1243..1290
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1299..1385
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1477..1504
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2007..2198
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2192..2214)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2300..2439
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2355..2374
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2454..2471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2625..3213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2705..2724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3384..4175,4176..4244))
                     /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     4007..4026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4245..4349)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4525..5403)
                     /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 tgctgccacc gctgagcaat aactagcata accccttggg
     1261 gcctctaaac gggtcttgag gggttttttg ccaggcatca aataaaacga aaggctcagt
     1321 cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1381 caaatccgcc gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag
     1441 gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc
     1501 tttttgcgtt tcagatctcg ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc
     1561 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
     1621 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
     1681 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
     1741 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
     1801 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
     1861 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
     1921 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
     1981 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
     2041 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
     2101 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
     2161 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2221 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2281 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     2341 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa
     2401 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     2461 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2521 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2581 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2641 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2701 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2761 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2821 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2881 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2941 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3001 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3061 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3121 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3181 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3241 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3301 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3361 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3421 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3481 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3541 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3601 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3661 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     3721 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3781 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3841 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3901 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3961 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     4021 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4081 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4141 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4201 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4261 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4321 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc accttcactc
     4381 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     4441 aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttgc
     4501 tcctagggtc tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc
     4561 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
     4621 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
     4681 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
     4741 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
     4801 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
     4861 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
     4921 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
     4981 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
     5041 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
     5101 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
     5161 gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg
     5221 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
     5281 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
     5341 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
     5401 cat
//
