LOCUS       62056_23215        7658 bp DNA     circular SYN 02-MAY-2020
DEFINITION  Cloning vector pZS2-200, complete sequence.
ACCESSION   MN744968
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7658)
  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 7658)
  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 7658)
  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..7658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      19..64
                     /label=RNAI
                     /note="RNAI"
                     /regulatory_class="terminator"
     regulatory      complement(65..238)
                     /label=TSAL2
                     /note="TSAL2"
                     /regulatory_class="terminator"
     CDS             complement(263..976)
                     /label=mCerulean
                     /note="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
     protein_bind    complement(1032..1048)
                     /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)."
     promoter        complement(1056..1086)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1101..1122)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     regulatory      1353..1852
                     /label=putative PdddW
                     /note="putative PdddW"
                     /regulatory_class="promoter"
     CDS             1889..2602
                     /label=mVenus
                     /note="Venus YFP with monomerizing A206K mutation (Nagai et
                     al., 2002; Kremers et al., 2006)"
     regulatory      2621..2743
                     /label=TR2-17
                     /note="TR2-17"
                     /regulatory_class="terminator"
     regulatory      2744..2820
                     /label=TL17
                     /note="TL17"
                     /regulatory_class="terminator"
     regulatory      2833..2885
                     /label=BS7
                     /note="BS7"
                     /regulatory_class="terminator"
     terminator      complement(2910..2937)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2953..3024)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3042..3749)
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     RBS             complement(3758..3778)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(3808..4307)
                     /label=putative PdmdA
                     /note="putative PdmdA"
                     /regulatory_class="promoter"
     CDS             4538..5329
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      5363..5457
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5949..6171
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6219..7166
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
ORIGIN
        1 gcggccgcaa aaggaaaaga tccggcaaac aaaccaccgt tggtagcggt ggtttttttg
       61 tttggatcga caatcttcgt aagcgtcatc aataagcgta aaaaaaccgg gcaatgcccg
      121 gttttttaat gagaaatttt acctgtcgta gccgccacca tccggcaaag aagcatacaa
      181 ggcttttggc ttatagctac gtagcgcatt gcgtcgcagc acaatcccgg caccgatcaa
      241 gtcttcgcga tgattattat tatttataca gctcatccat gccatgcgtg atgccagcag
      301 ccgttacaaa ttccagcagc accatgtgat cgcgcttctc attaggatct ttgctcagct
      361 tagattgagt gctcagatag tgattgtccg gcagcagaac aggaccgtcg ccaatcgggg
      421 tattttgttg ataatggtca gccagttgca cagagccgtc ctcgatatta tggcgaattt
      481 taaaatgcgc cttaatacca ttcttttgtt tatcggcggt gatatacaca ttatcgctaa
      541 tcgcattata ttccagttta tggcccagga tattaccatc ttccttaaaa tcgatgccct
      601 tcagctcaat acgattcacc agagtgtcgc cttcaaactt cacctctgcg cgagtcttgt
      661 aattaccatc atctttaaag aaaatggtgc gctcctgaac gtaaccctct ggcatggcgc
      721 tcttaaaaaa atcgtgctgc ttcatgtggt ctggatagcg agcgaagcat tgaacacccc
      781 aggtcagggt ggtaaccagg gtaggccaag gcaccggcag cttacccgtc gtacaaatga
      841 acttcagggt cagcttaccg taggttgcgt cgccttcacc ctcgccgcta acgctgaact
      901 tatgaccgtt gacatcaccg tccagttcga ccaggatagg aaccacacca gtaaacagct
      961 cctcgccctt gctcattttt ttttcctcct tattttctcc aggaagatct tcagctgttt
     1021 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1081 tgtaaagcct ggggtgccta atgagtgagc taactcacat tgtcgacaaa aataatgaga
     1141 atcaatagaa cttccgagaa gttcagccgc taataatcgc cctgctccat tgtgcgccgc
     1201 aataaaagta ccggcattac gggtgcattg gcgcgccaaa tgcggccatc tgtgggcaac
     1261 ctgtgcggta agacccaaac ttagtgtaat aggtatccta tgattatttt ttcatttgat
     1321 gccaaaaaga caatgaaccc ccgctcgagc gggggtcgcc tcaccatact cctgacttat
     1381 gatccggatg gtcacatcct cgaaagcagc ctccagttcc cccagctgcg gaatcagcca
     1441 aaaggcgaca aagcccgacg gcgcagtcac ggtcaccacc ttgccctgcg ccgcttgccg
     1501 gatttgggca accgccgcct cgatctggcc gaacccacct tgcaaagccc gcagcaggtc
     1561 ccggcctgac tgggtcatac gcaccggttt gtgactgcga tcgaacagcg gactgcccaa
     1621 atccgcctcc agcatcgcga tccggcgcga gaccgcgggt tgcgacacgt tcagctctgc
     1681 cgctgccgcg gtcagcgtgc cgcgacgcgc ggccgcgtag aatgcagtca agccggagaa
     1741 gcttggcaaa cgctccatat cataacctca tcgtataatc tgatgcagtt ttacgcgatt
     1801 gcgccccccc tgcaagagcg actaactgac cccaagcaca aaggagccca tccgggatcc
     1861 cggtgcagaa aataaggagg aaaaaaaaat gagcaaaggt gaagaactgt tcaccggcgt
     1921 tgtgccaatt ctggttgagc tggatggtga cgtgaatggc cacaaatttt ccgtgtctgg
     1981 tgaaggcgag ggtgatgcta cttatggcaa actgactctg aaactgatct gtaccaccgg
     2041 caaactgcct gttccgtggc caactctggt cactactctg ggttacggcc tgatgtgttt
     2101 tgcgcgttac ccggatcaca tgaaacagca tgacttcttc aaatctgcca tgccggaagg
     2161 ctatgtccaa gaacgtacga tctttttcaa ggacgacggc aactataaaa cccgtgccga
     2221 agttaaattc gagggtgaca ccctggttaa ccgcatcgaa ctgaaaggca ttgacttcaa
     2281 agaggacggc aacattctgg gtcacaagct ggaatacaac tacaactccc acaacgttta
     2341 cattactgct gacaagcaga aaaacggcat caaagcaaac ttcaagatcc gtcacaacat
     2401 tgaagatggt ggcgtacagc tggcagatca ctaccagcag aacactccaa tcggtgatgg
     2461 cccagtactg ctgccagata accattacct gtcctaccag agcaaactgt ctaaagaccc
     2521 gaacgaaaaa cgtgaccaca tggtactgct ggaatttgtt accgcggcag gcattaccca
     2581 cggtatggac gaactgtata aataatgcag gtcgtctcgg atcgagaagg acacggttaa
     2641 tactaggcct gctggctggt aatcgccagc aggccttttt atttggggga gagggaagtc
     2701 atgaaaaaac taacctttga aattcgatct ccaccacatc agctctgaag caacgtaaaa
     2761 aaacccgccc cggcgggttt ttttataccc gtagtatccc cacttatcta caatagctgt
     2821 ccttaattaa ggttgaataa ataaaaacag ccgttgccag aaagaggcac ggctgttttt
     2881 attttctagt gagaccggga gcagttaaac gcagaaaggc ccacccgaag gtgagccagt
     2941 gtgactctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     3001 cgactgagcc tttcgtttta tttgatgcct ggttattatt atttgtacag ctcatccatg
     3061 ccaccggtag aatgacgacc ctccgcgcgc tcatattgct ctacgatcgt ataatcttca
     3121 ttatgagagg tgatgtccag tttaatattc acattgtacg cgccaggcag ctgcacaggt
     3181 ttcttggctt tgtacgtggt tttcacttca gcgtcataat ggccgccatc tttcagtttc
     3241 aggcgctgtt taatttcgcc tttcagcgca ccatcttccg gatacatgcg ttcgctagac
     3301 gcctcccaac ccatcgtctt tttctgcatc accgggccat cagatggaaa attagtacca
     3361 cgcagtttaa ctttatagat gaactcgcca tcctgcaggg aggagtcctg agtgacggtc
     3421 acgacaccac catcttcaaa attcattacg cgttcccact tgaaaccttc cggaaaagac
     3481 agcttcagat agtccgggat atccgctggg tgtttaacat acgctttaga accgtacata
     3541 aattgcgggc tcaggatgtc ccacgcaaaa ggcagcgggc cgcctttagt cactttcagt
     3601 ttggcggtct gggtgccttc atacggacgg ccctcgcctt cgccttcgat ctcgaactcg
     3661 tgaccgttaa cagaaccctc catgtgaact ttgaagcgca tgaactcttt aatgatagcc
     3721 atgttatcct cctcgccctt ggaaaccatg gcaggtgctc cttcttaaag ttaaacaaaa
     3781 ttatttctag attttctcaa gcctagggat cggatctcca gacgagcgtt atggtatgcg
     3841 ggccgaacga gaggcccggc ggtgatgcgg cggtgcaatc gcccttccgg cattcgctgc
     3901 gaccgtcatt gcggctcttg ccctcggacg actcacgtca tagggtctta aatgtcagac
     3961 ttttaaatgt cagacaaatg aattcttgcc ggcgccgcgc cggtgcgacc cggcgtcagg
     4021 accacagcca tgcagatcga ccccaacagc gccaccgatc ttccggtgca gatcgcccag
     4081 gcgatcaagg acgcgatcgt ttcgggcgcg ttgatcgtgg atgaacgcct gccgtccgag
     4141 gctgaactgg ccgaccagtt caaggtctcg cgccccacgg tgcgcgaggc gctcaagcgg
     4201 ctggcggcgc aatcgctgat ccgcacccag cgcggcgcct ttggcggcgc tttcgtgaac
     4261 cggctgagtt tcgaggatgc gcaggcccag cagatcacca cctcgacttc cccccggggg
     4321 gatatcaaca ggagtccaag cgaccggtgg ttgcatgtct agctagctag aacaggacta
     4381 gctaatggtt tcttagacgt cggaattgcc agctggggcg ccctctggta aggttgggaa
     4441 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     4501 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg agagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     5401 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
     5461 agcactaggg acagtaagac gggtaagcct gttgatgata ccgctgcctt actgggtgca
     5521 ttagccagtc tgaatgacct gtcacgggat aatccgaagt ggtcagactg gaaaatcaga
     5581 gggcaggaac tgctgaacag caaaaagtca gatagcacca catagcagac ccgccataaa
     5641 acgccctgag aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat
     5701 ccataaaagg cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc
     5761 gaactgaatg tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa
     5821 tattcagcga tttgcccgag cttgcgaggg tgctacttaa gcctttaggg ttttaaggtc
     5881 tgttttgtag aggagcaaac agcgtttgcg acatcctttt gtaatactgc ggaactgact
     5941 aaagtagtga gttatacaca gggctgggat ctattctttt tatctttttt tattctttct
     6001 ttattctata aattataacc acttgaatat aaacaaaaaa aacacacaaa ggtctagcgg
     6061 aatttacaga gggtctagca gaatttacaa gttttccagc aaaggtctag cagaatttac
     6121 agatacccac aactcaaagg aaaaggacta gtaattatca ttgactagcc catctcaatt
     6181 ggtatagtga ttaaaatcac ctagaccaat tgagatgtat gtctgaatta gttgttttca
     6241 aagcaaatga actagcgatt agtcgctatg acttaacgga gcatgaaacc aagctaattt
     6301 tatgctgtgt ggcactactc aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg
     6361 tatcgttcac ttataaccaa tacgctcaga tgatgaacat cagtagggaa aatgcttatg
     6421 gtgtattagc taaagcaacc agagagctga tgacgagaac tgtggaaatc aggaatcctt
     6481 tggttaaagg ctttgagatt ttccagtgga caaactatgc caagttctca agcgaaaaat
     6541 tagaattagt ttttagtgaa gagatattgc cttatctttt ccagttaaaa aaattcataa
     6601 aatataatct ggaacatgtt aagtcttttg aaaacaaata ctctatgagg atttatgagt
     6661 ggttattaaa agaactaaca caaaagaaaa ctcacaaggc aaatatagag attagccttg
     6721 atgaatttaa gttcatgtta atgcttgaaa ataactacca tgagtttaaa aggcttaacc
     6781 aatgggtttt gaaaccaata agtaaagatt taaacactta cagcaatatg aaattggtgg
     6841 ttgataagcg aggccgcccg actgatacgt tgattttcca agttgaacta gatagacaaa
     6901 tggatctcgt aaccgaactt gagaacaacc agataaaaat gaatggtgac aaaataccaa
     6961 caaccattac atcagattcc tacctacgta acggactaag aaaaacacta cacgatgctt
     7021 taactgcaaa aattcagctc accagttttg aggcaaaatt tttgagtgac atgcaaagta
     7081 agcatgatct caatggttcg ttctcatggc tcacgcaaaa acaacgaacc acactagaga
     7141 acatactggc taaatacgga aggatctgag gttcttatgg ctcttgtatc tatcagtgaa
     7201 gcatcaagac taacaaacaa aagtagaaca actgttcacc gttagatatc aaagggaaaa
     7261 ctgtccatat gcacagatga aaacggtgta aaaaagatag atacatcaga gcttttacga
     7321 gtttttggtg catttaaagc tgttcaccat gaacagatcg acaatgtaac agatgaacag
     7381 catgtaacac ctaatagaac aggtgaaacc agtaaaacaa agcaactaga acatgaaatt
     7441 gaacacctga gacaacttgt tacagctcaa cagtcacaca tagacagcct gaaacaggcg
     7501 atgctgctta tcgaatcaaa gctgccgaca acacgggagc cagtgacgcc tcccgtgggg
     7561 aaaaaatcat ggcaattctg gaagaaatag cgctttcagc cggcaaacct gaagccggat
     7621 ctgcgattct gataacaaac tagcaacacc agaacagc
//
