LOCUS       sequence-244262-        5618 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 5618)
  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 5618)
  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_v5
FEATURES             Location/Qualifiers
     source          1..5618
                     /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"
     CDS             complement(1766..2716)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2764..2987
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3479..3573)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3599..4390,4391..4459))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDKRNRQIAEIGAS
                     LIKHW"
     primer_bind     4222..4241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4460..4564)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4740..5618)
                     /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 aactagcgta cgggttttgc
     1261 tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt
     1321 cagccggttt gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac
     1381 gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg
     1441 tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc
     1501 atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc
     1561 atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa
     1621 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc
     1681 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     1741 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     1801 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     1861 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     1921 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     1981 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     2041 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     2101 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     2161 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     2221 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     2281 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     2341 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     2401 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     2461 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     2521 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     2581 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     2641 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     2701 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     2761 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
     2821 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     2881 gacctttgtg tgtttttttt tgtttatatt caagtggtta taatttatag aataaagaaa
     2941 gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact actttagtca
     3001 gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct
     3061 taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct
     3121 tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc
     3181 gctcacggct ctggcagtga atgggggtaa atggcactac aggctccttt tatggattca
     3241 tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat
     3301 ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga
     3361 ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac
     3421 ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta gtgcttggat
     3481 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     3541 ctgaggtcat tactggatct atcaacagga gtccaagcgc gtaaacttgg tctgacagtt
     3601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt ttatccatag
     3661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4561 cgcgcacatt tccccgaaaa gtgccacctt cactctagat ttcagtgcaa tttatctctt
     4621 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat
     4681 gccccgcgcc caccggaagg agctgactgg gttgctccta gggtctgatt cgttaccaat
     4741 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
     4801 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
     4861 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
     4921 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
     4981 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
     5041 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
     5101 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
     5161 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
     5221 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
     5281 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
     5341 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
     5401 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
     5461 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
     5521 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
     5581 tatcccggca gcaggggatc attttgcgct tcagccat
//
