LOCUS       62056_6250        5970 bp DNA     circular SYN 02-MAY-2020
DEFINITION  Cloning vector pCG303, complete sequence.
ACCESSION   MN744964
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5970)
  AUTHORS   Gao C, Fernandez VI, Lee KS, Fenizia S, Pohnert G, Seymour JR, Raina
            J-B., Stocker R.
  TITLE     Single-cell bacterial transcription measurements reveal the 
            importance of dimethylsulfoniopropionate (DMSP) hotspots in ocean 
            sulfur cycling
  JOURNAL   Nat Commun 11 (1) (2020) In press
REFERENCE   2  (bases 1 to 5970)
  AUTHORS   Gao C.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2019) Biological Engineering, Massachusetts 
            Institute of Technology, 15 Vassar Street, Cambridge, MA 20139, USA
REFERENCE   3  (bases 1 to 5970)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            11 (1) (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-NOV-2019) Biological Engineering, Massachusetts Institute of 
            Technology, 15 Vassar Street, Cambridge, MA 20139, USA"
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     regulatory      2887..3386
                     /label=putative PdddW
                     /note="putative PdddW"
                     /regulatory_class="promoter"
     CDS             3387..4082
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     regulatory      4107..4229
                     /label=TR2-17
                     /note="TR2-17"
                     /regulatory_class="terminator"
     regulatory      4230..4306
                     /label=TL17
                     /note="TL17"
                     /regulatory_class="terminator"
     regulatory      4319..4371
                     /label=BS7
                     /note="BS7"
                     /regulatory_class="terminator"
     terminator      complement(4396..4423)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4439..4510)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4722..5513)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc
     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac
     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     2641 ggcagaataa atgatgtcga caaaaataat gagaatcaat agaacttccg agaagttcag
     2701 ccgctaataa tcgccctgct ccattgtgcg ccgcaataaa agtaccggca ttacgggtgc
     2761 attggcgcgc caaatgcggc catctgtggg caacctgtgc ggtaagaccc aaacttagtg
     2821 taataggtat cctatgatta ttttttcatt tgatgccaaa aagacaatga acccccgctc
     2881 gagcgggggt cgcctcacca tactcctgac ttatgatccg gatggtcaca tcctcgaaag
     2941 cagcctccag ttcccccagc tgcggaatca gccaaaaggc gacaaagccc gacggcgcag
     3001 tcacggtcac caccttgccc tgcgccgctt gccggatttg ggcaaccgcc gcctcgatct
     3061 ggccgaaccc accttgcaaa gcccgcagca ggtcccggcc tgactgggtc atacgcaccg
     3121 gtttgtgact gcgatcgaac agcggactgc ccaaatccgc ctccagcatc gcgatccggc
     3181 gcgagaccgc gggttgcgac acgttcagct ctgccgctgc cgcggtcagc gtgccgcgac
     3241 gcgcggccgc gtagaatgca gtcaagccgg agaagcttgg caaacgctcc atatcataac
     3301 ctcatcgtat aatctgatgc agttttacgc gattgcgccc cccctgcaag agcgactaac
     3361 tgaccccaag cacaaaggag cccatcatgg tgagcgagct gattaaggag aacatgcaca
     3421 tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg
     3481 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggcggtcgag ggcggccctc
     3541 tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaaa accttcatca
     3601 accacaccca gggcatcccc gacttcttta agcagtcctt ccccgagggc ttcacatggg
     3661 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     3721 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc
     3781 ctgtgatgca gaagaaaaca ctcggctggg aggcctccac cgagaccctg taccccgctg
     3841 acggcggcct ggaaggcaga gccgacatgg ccctgaagct cgtgggcggg ggccacctga
     3901 tctgcaactt gaagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg
     3961 gcgtctacta tgtggacaga agactggaaa gaatcaagga ggccgacaaa gagacctacg
     4021 tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca
     4081 gataataata atgcaggtcg tctcggatcg agaaggacac ggttaatact aggcctgctg
     4141 gctggtaatc gccagcaggc ctttttattt gggggagagg gaagtcatga aaaaactaac
     4201 ctttgaaatt cgatctccac cacatcagct ctgaagcaac gtaaaaaaac ccgccccggc
     4261 gggttttttt atacccgtag tatccccact tatctacaat agctgtcctt aattaaggtt
     4321 gaataaataa aaacagccgt tgccagaaag aggcacggct gtttttattt tctagtgaga
     4381 ccgggagcag ttaaacgcag aaaggcccac ccgaaggtga gccagtgtga ctctagtaga
     4441 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     4501 gttttatttg atgcctggtc gccttgcgta taatatttgc ccatgggggt gggcgaagaa
     4561 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     4621 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     4681 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     4741 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     4801 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     4861 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     4921 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     4981 atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     5041 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     5101 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     5161 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     5221 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     5281 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca
     5341 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     5401 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     5461 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     5521 gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc
     5581 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
     5641 ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc
     5701 ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag
     5761 tagctgacat tcatcccagg tggcactttt cggggaaatg tgcgcgcccg cgttcctgct
     5821 ggcgctgggc ctgtttctgg cgctggactt cccgctgttc cgtcagcagc ttttcgccca
     5881 cggccttgat gatcgcggcg gccttggcct gcatatcccg attcaacggc cccagggcgt
     5941 ccagaacggg cttcaggcgc tcccgaaggt
//
