LOCUS       62056_6240        7974 bp DNA     circular SYN 02-MAY-2020
DEFINITION  Cloning vector pCG301, complete sequence.
ACCESSION   MN744962
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7974)
  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 7974)
  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 7974)
  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..7974
                     /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      2656..2701
                     /label=RNAI
                     /note="RNAI"
                     /regulatory_class="terminator"
     regulatory      complement(2702..2875)
                     /label=TSAL2
                     /note="TSAL2"
                     /regulatory_class="terminator"
     CDS             complement(2900..3607)
                     /label=mTFP1
                     /note="monomeric teal (cyan) fluorescent protein (Ai et
                     al., 2006)"
     regulatory      complement(3608..3829)
                     /label=putative PdmdA
                     /note="putative PdmdA"
                     /regulatory_class="promoter"
     regulatory      4061..4560
                     /label=putative PdddW
                     /note="putative PdddW"
                     /regulatory_class="promoter"
     CDS             4561..5256
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     regulatory      5281..5403
                     /label=TR2-17
                     /note="TR2-17"
                     /regulatory_class="terminator"
     regulatory      5404..5480
                     /label=TL17
                     /note="TL17"
                     /regulatory_class="terminator"
     regulatory      5493..5545
                     /label=BS7
                     /note="BS7"
                     /regulatory_class="terminator"
     terminator      complement(5570..5597)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5613..5684)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(5702..6415)
                     /label=mVenus
                     /note="Venus YFP with monomerizing A206K mutation (Nagai et
                     al., 2002; Kremers et al., 2006)"
     protein_bind    complement(6485..6501)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             complement(6726..7517)
                     /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 atgatgatcc ggcaaacaaa ccaccgttgg tagcggtggt ttttttgttt
     2701 ggatcgacaa tcttcgtaag cgtcatcaat aagcgtaaaa aaaccgggca atgcccggtt
     2761 ttttaatgag aaattttacc tgtcgtagcc gccaccatcc ggcaaagaag catacaaggc
     2821 ttttggctta tagctacgta gcgcattgcg tcgcagcaca atcccggcac cgatcaagtc
     2881 ttcgcgatga ttattattac ttgtacagct cgtccatgcc gtcggtggag ttgcgggcca
     2941 cggcgctctc gtaaacggtc accttgttgt agtccttgtc gtggttcagg atctcgatgc
     3001 ggtggtccac aaagtgatag tcgggcagct tcaccgcctt cttggccctg tagatggtct
     3061 tgaagtcaac gcggtggtgg ccgccgccct ccagcagcag cttgtgcttg acgtcgccct
     3121 tcagcacgcc gtcgcgcacg tacatcctct cggtggaggc gtcccagccg gtggtcttct
     3181 tctgcatcac ggggccgttg ggggggaagt tctcgccctt gaggtgtatc tcgtagatga
     3241 aggagtcctc ctccatggag atgtcggact tcaccttcac gatgcccttg tcctcgaagg
     3301 tcatggtgcg ctcccaagag tagccctcgg ggaaggactg cttgaagtag ttggggatgt
     3361 cgtcggggta cttggtgaag gccctgttgc cgtaggcgaa cgcggtggtc agaatgtcgt
     3421 aggagaaggg caggggggct ccctccttca cctccaggtt gatggtgttg gtgccgtcgt
     3481 agggcttgcc ctcgccctcg ccctcgatca cgaaggcgtg gccattcacg ttgccctcca
     3541 tcttcagctt gatcttcatg tcgggcttga ttacgcccat tgtggtctcc tcgcccttgc
     3601 tcaccatgat cggatctcca gacgagcgtt atggtatgcg ggccgaacga gaggcccggc
     3661 ggtgatgcgg cggtgcaatc gcccttccgg cattcgctgc gaccgtcatt gcggctcttg
     3721 ccctcggacg actcacgtca tagggtctta aatgtcagac ttttaaatgt cagacaaatg
     3781 aattcttgcc ggcgccgcgc cggtgcgacc cggcgtcagg accacagccg tcgacaaaaa
     3841 taatgagaat caatagaact tccgagaagt tcagccgcta ataatcgccc tgctccattg
     3901 tgcgccgcaa taaaagtacc ggcattacgg gtgcattggc gcgccaaatg cggccatctg
     3961 tgggcaacct gtgcggtaag acccaaactt agtgtaatag gtatcctatg attatttttt
     4021 catttgatgc caaaaagaca atgaaccccc gctcgagcgg gggtcgcctc accatactcc
     4081 tgacttatga tccggatggt cacatcctcg aaagcagcct ccagttcccc cagctgcgga
     4141 atcagccaaa aggcgacaaa gcccgacggc gcagtcacgg tcaccacctt gccctgcgcc
     4201 gcttgccgga tttgggcaac cgccgcctcg atctggccga acccaccttg caaagcccgc
     4261 agcaggtccc ggcctgactg ggtcatacgc accggtttgt gactgcgatc gaacagcgga
     4321 ctgcccaaat ccgcctccag catcgcgatc cggcgcgaga ccgcgggttg cgacacgttc
     4381 agctctgccg ctgccgcggt cagcgtgccg cgacgcgcgg ccgcgtagaa tgcagtcaag
     4441 ccggagaagc ttggcaaacg ctccatatca taacctcatc gtataatctg atgcagtttt
     4501 acgcgattgc gccccccctg caagagcgac taactgaccc caagcacaaa ggagcccatc
     4561 atggtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg
     4621 aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
     4681 accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga catcctggct
     4741 accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat ccccgacttc
     4801 tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata cgaagacggg
     4861 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
     4921 aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc
     4981 tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg cagagccgac
     5041 atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac cacatacaga
     5101 tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga cagaagactg
     5161 gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt ggctgtggcc
     5221 agatactgcg acctccctag caaactgggg cacagataat aataatgcag gtcgtctcgg
     5281 atcgagaagg acacggttaa tactaggcct gctggctggt aatcgccagc aggccttttt
     5341 atttggggga gagggaagtc atgaaaaaac taacctttga aattcgatct ccaccacatc
     5401 agctctgaag caacgtaaaa aaacccgccc cggcgggttt ttttataccc gtagtatccc
     5461 cacttatcta caatagctgt ccttaattaa ggttgaataa ataaaaacag ccgttgccag
     5521 aaagaggcac ggctgttttt attttctagt gagaccggga gcagttaaac gcagaaaggc
     5581 ccacccgaag gtgagccagt gtgactctag tagagagcgt tcaccgacaa acaacagata
     5641 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggttattatt
     5701 atttatacag ttcgtccata ccgtgggtaa tgcctgccgc ggtaacaaat tccagcagta
     5761 ccatgtggtc acgtttttcg ttcgggtctt tagacagttt gctctggtag gacaggtaat
     5821 ggttatctgg cagcagtact gggccatcac cgattggagt gttctgctgg tagtgatctg
     5881 ccagctgtac gccaccatct tcaatgttgt gacggatctt gaagtttgct ttgatgccgt
     5941 ttttctgctt gtcagcagta atgtaaacgt tgtgggagtt gtagttgtat tccagcttgt
     6001 gacccagaat gttgccgtcc tctttgaagt caatgccttt cagttcgatg cggttaacca
     6061 gggtgtcacc ctcgaattta acttcggcac gggttttata gttgccgtcg tccttgaaaa
     6121 agatcgtacg ttcttggaca tagccttccg gcatggcaga tttgaagaag tcatgctgtt
     6181 tcatgtgatc cgggtaacgc gcaaaacaca tcaggccgta acccagagta gtgaccagag
     6241 ttggccacgg aacaggcagt ttgccggtgg tacagatcag tttcagagtc agtttgccat
     6301 aagtagcatc accctcgcct tcaccagaca cggaaaattt gtggccattc acgtcaccat
     6361 ccagctcaac cagaattggc acaacgccgg tgaacagttc ttcacctttg ctcatgctta
     6421 atttctcctc tttaattcta gatgtgtgaa attgttatcc gctcacaatt gaatctaagt
     6481 atcattgtta tccgctcaca agtcaacact ctttttgata aatcgccttg cgtataatat
     6541 ttgcccatgg gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     6601 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     6661 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     6721 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     6781 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     6841 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     6901 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     6961 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     7021 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     7081 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     7141 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     7201 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     7261 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     7321 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     7381 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     7441 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     7501 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     7561 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     7621 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc
     7681 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg
     7741 ttttcccttg tccagatagc ccagtagctg acattcatcc caggtggcac ttttcgggga
     7801 aatgtgcgcg cccgcgttcc tgctggcgct gggcctgttt ctggcgctgg acttcccgct
     7861 gttccgtcag cagcttttcg cccacggcct tgatgatcgc ggcggccttg gcctgcatat
     7921 cccgattcaa cggccccagg gcgtccagaa cgggcttcag gcgctcccga aggt
//
