LOCUS       62056_6255        7974 bp DNA     circular SYN 02-MAY-2020
DEFINITION  Cloning vector pCG401, complete sequence.
ACCESSION   MN744965
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..3595)
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     regulatory      complement(3596..3817)
                     /label=putative PdmdA
                     /note="putative PdmdA"
                     /regulatory_class="promoter"
     regulatory      4049..4548
                     /label=putative PdddW
                     /note="putative PdddW"
                     /regulatory_class="promoter"
     CDS             4549..5256
                     /label=mTFP1
                     /note="monomeric teal (cyan) fluorescent protein (Ai et
                     al., 2006)"
     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 ttattattat ctgtgcccca gtttgctagg gaggtcgcag tatctggcca
     2941 cagccacctc gtgctgctcg acgtaggtct ctttgtcggc ctccttgatt ctttccagtc
     3001 ttctgtccac atagtagacg ccgggcatct tgaggttctt agcgggtttc ttggatctgt
     3061 atgtggtctt caagttgcag atcaggtggc ccccgcccac gagcttcagg gccatgtcgg
     3121 ctctgccttc caggccgccg tcagcggggt acagggtctc ggtggaggcc tcccagccga
     3181 gtgttttctt ctgcatcaca gggccgttgg atgggaagtt cacccctctg atcttgacgt
     3241 tgtagatgag gcagccgtcc tggaggctgg tgtcctgggt agcggtcagc acgcccccgt
     3301 cttcgtatgt ggtgactctc tcccatgtga agccctcggg gaaggactgc ttaaagaagt
     3361 cggggatgcc ctgggtgtgg ttgatgaagg ttttgctgcc gtacatgaag ctggtagcca
     3421 ggatgtcgaa ggcgaagggg agagggccgc cctcgaccgc cttgattctc atggtctggg
     3481 tgccctcgta gggcttgcct tcgccctcgg atgtgcactt gaagtggtgg ttgttcacgg
     3541 tgccctccat gtacagcttc atgtgcatgt tctccttaat cagctcgctc accatgatcg
     3601 gatctccaga cgagcgttat ggtatgcggg ccgaacgaga ggcccggcgg tgatgcggcg
     3661 gtgcaatcgc ccttccggca ttcgctgcga ccgtcattgc ggctcttgcc ctcggacgac
     3721 tcacgtcata gggtcttaaa tgtcagactt ttaaatgtca gacaaatgaa ttcttgccgg
     3781 cgccgcgccg gtgcgacccg gcgtcaggac cacagccgtc gacaaaaata atgagaatca
     3841 atagaacttc cgagaagttc agccgctaat aatcgccctg ctccattgtg cgccgcaata
     3901 aaagtaccgg cattacgggt gcattggcgc gccaaatgcg gccatctgtg ggcaacctgt
     3961 gcggtaagac ccaaacttag tgtaataggt atcctatgat tattttttca tttgatgcca
     4021 aaaagacaat gaacccccgc tcgagcgggg gtcgcctcac catactcctg acttatgatc
     4081 cggatggtca catcctcgaa agcagcctcc agttccccca gctgcggaat cagccaaaag
     4141 gcgacaaagc ccgacggcgc agtcacggtc accaccttgc cctgcgccgc ttgccggatt
     4201 tgggcaaccg ccgcctcgat ctggccgaac ccaccttgca aagcccgcag caggtcccgg
     4261 cctgactggg tcatacgcac cggtttgtga ctgcgatcga acagcggact gcccaaatcc
     4321 gcctccagca tcgcgatccg gcgcgagacc gcgggttgcg acacgttcag ctctgccgct
     4381 gccgcggtca gcgtgccgcg acgcgcggcc gcgtagaatg cagtcaagcc ggagaagctt
     4441 ggcaaacgct ccatatcata acctcatcgt ataatctgat gcagttttac gcgattgcgc
     4501 cccccctgca agagcgacta actgacccca agcacaaagg agcccatcat ggtgagcaag
     4561 ggcgaggaga ccacaatggg cgtaatcaag cccgacatga agatcaagct gaagatggag
     4621 ggcaacgtga atggccacgc cttcgtgatc gagggcgagg gcgagggcaa gccctacgac
     4681 ggcaccaaca ccatcaacct ggaggtgaag gagggagccc ccctgccctt ctcctacgac
     4741 attctgacca ccgcgttcgc ctacggcaac agggccttca ccaagtaccc cgacgacatc
     4801 cccaactact tcaagcagtc cttccccgag ggctactctt gggagcgcac catgaccttc
     4861 gaggacaagg gcatcgtgaa ggtgaagtcc gacatctcca tggaggagga ctccttcatc
     4921 tacgagatac acctcaaggg cgagaacttc ccccccaacg gccccgtgat gcagaagaag
     4981 accaccggct gggacgcctc caccgagagg atgtacgtgc gcgacggcgt gctgaagggc
     5041 gacgtcaagc acaagctgct gctggagggc ggcggccacc accgcgttga cttcaagacc
     5101 atctacaggg ccaagaaggc ggtgaagctg cccgactatc actttgtgga ccaccgcatc
     5161 gagatcctga accacgacaa ggactacaac aaggtgaccg tttacgagag cgccgtggcc
     5221 cgcaactcca ccgacggcat ggacgagctg tacaagtaat 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
//
