LOCUS       62056_18825        8792 bp DNA     circular SYN 28-FEB-2021
DEFINITION  Cloning vector pRGC-BHR, complete sequence.
ACCESSION   MW148405
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8792)
  AUTHORS   Mellini M, Lucidi M, Imperi F, Visca P, Leoni L, Rampioni G.
  TITLE     Generation of genetic tools for gauging multiple gene expression at 
            single cell level
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8792)
  AUTHORS   Mellini M, Lucidi M, Imperi F, Visca P, Leoni L, Rampioni G.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2020) Science, University Roma Tre, Viale 
            Guglielmo Marconi 446, Roma 00146, Italia
REFERENCE   3  (bases 1 to 8792)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-OCT-2020) Science, University Roma Tre, Viale Guglielmo Marconi 
            446, Roma 00146, Italia"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(528..722)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(791..1861)
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     gene            complement(2054..2269)
                     /gene="parG"
                     /label=parG
     CDS             complement(2054..2269)
                     /codon_start=1
                     /transl_table=11
                     /gene="parG"
                     /product="ParG"
                     /label=parG
                     /note="pVS1 partitioning protein"
                     /protein_id="QRZ20922.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     CDS             complement(2293..2919)
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             complement(3006..3221)
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /protein_id="QRZ20924.1"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             complement(3218..3904)
                     /codon_start=1
                     /transl_table=11
                     /product="resolvase"
                     /label=resolvase
                     /note="derived from pVS1"
                     /protein_id="QRZ20925.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             complement(4161..4691)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(4880..4908)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(4926..5030)
                     /label=AmpR promoter
     primer_bind     5269..5285
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5295..5313
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5331..6047)
                     /label=ECFP
                     /note="enhanced CFP"
     regulatory      complement(6054..6065)
                     /label=strong bacterial ribosome binding site
                     /note="strong bacterial ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
     primer_bind     6083..6099
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      6120..6131
                     /label=strong bacterial ribosome binding site
                     /note="strong bacterial ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
     CDS             6138..6851
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     terminator      6874..6968
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(7019..7035)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7058..7069
                     /label=strong bacterial ribosome binding site
                     /note="strong bacterial ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
     CDS             7076..7783
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     promoter        complement(7806..7824)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      complement(8016..8604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcatacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
       61 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      121 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      181 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      241 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgccct
      301 ggtatgagtc agcaacacct tcttcacgag gcagacctca gcgccggcgg tcgagtggcg
      361 acggcgcggc ttgtccgcgc cctggtagat tgcctggccg taggccagcc atttttgagc
      421 ggccagcggc cgcgataggc cgacgcgaag cggcggggcg tagggagcgc agcgaccgaa
      481 gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc agacagttat gcacaggcca
      541 ggcgggtttt aagagtttta ataagtttta aagagtttta ggcggaaaaa tcgccttttt
      601 tctcttttat atcagtcact tacatgtgtg accggttccc aatgtacggc tttgggttcc
      661 caatgtacgg gttccggttc ccaatgtacg gctttgggtt cccaatgtac gtgctatcca
      721 caggaaagag accttttcga cctttttccc ctgctagggc aatttgccct agcatctgct
      781 ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag gttgcggtag cgcatgacta
      841 ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta ctccggcagg tcatttgacc
      901 cgatcagctt gcgcacggtg aaacagaact tcttgaactc tccggcgctg ccactgcgtt
      961 cgtagatcgt cttgaacaac catctggctt ctgccttgcc tgcggcgcgg cgtgccaggc
     1021 ggtagagaaa acggccgatg ccgggatcga tcaaaaagta atcggggtga accgtcagca
     1081 cgtccgggtt cttgccttct gtgatctcgc ggtacatcca atcagctagc tcgatctcga
     1141 tgtactccgg ccgcccggtt tcgctcttta cgatcttgta gcggctaatc aaggcttcac
     1201 cctcggatac cgtcaccagg cggccgttct tggccttctt cgtacgctgc atggcaacgt
     1261 gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc tttctgcttt ccgccatcgg
     1321 ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg gaacacgcgg ccgggcttgt
     1381 ctcccttccc ttcccggtat cggttcatgg attcggttag atgggaaacc gccatcagta
     1441 ccaggtcgta atcccacaca ctggccatgc cggccggccc tgcggaaacc tctacgtgcc
     1501 cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg gtcacgcttc gacagacgga
     1561 aaacggccac gtccatgatg ctgcgactat cgcgggtgcc cacgtcatag agcatcggaa
     1621 cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct aatcgacggc gcaccggctg
     1681 ccggcggttg ccgggattct ttgcggattc gatcagcggc cgcttgccac gattcaccgg
     1741 ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg cgcggccttc aacttctcca
     1801 ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc ggatggtttg cgaccgctca
     1861 cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga caacccagcc
     1921 gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg cgtcggtgcc
     1981 tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc tactcattta
     2041 ttcatttgct catttactct ggtagctgcg cgatgtattc agatagcagc tcggtaatgg
     2101 tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc ggcaactgaa
     2161 agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca gccttgctgc
     2221 tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt ttctctttac
     2281 ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga cctcgcgggc
     2341 agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag tgcctgggta
     2401 gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg ccagcgcctc
     2461 ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa aggccgcttg
     2521 tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt cggcggtggc
     2581 ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg ccttgatcgc
     2641 ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt cgcgccggcc
     2701 gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg ttagcggttg
     2761 atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat cgactaacag
     2821 aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttgcgatgg tcgtcttgcc
     2881 tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt gcgtatttgt
     2941 ttatttactc atcgcatcat atacgcagcg accgcatgac gcaagctgtt ttactcaaat
     3001 acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct ggccggccag
     3061 gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac ggttgagacg
     3121 tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct ccgcctcgat ctcttcggta
     3181 atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc tcttggcgtt
     3241 cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg aaggcaccgc
     3301 gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg tacagggtcg
     3361 agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc tggtcgatca
     3421 gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca aacttcacgc
     3481 ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg gaacgctcga
     3541 actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg gtggtgtcgg
     3601 cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc tcggcaatgt
     3661 ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc acaacgtcgc
     3721 cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg gtgccggtga
     3781 tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt tggttggtca
     3841 agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg gcgctcttgt
     3901 tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga ctaaaacacg
     3961 cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact taggacttgt
     4021 gcgacatgat gcatcagcag caacagcagc aggggctgca tacttggtct gacaatcgat
     4081 gcgaattggc cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc
     4141 tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct
     4201 tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc
     4261 acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg
     4321 gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca
     4381 ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct
     4441 tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg
     4501 gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca
     4561 agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc
     4621 atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     4681 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     4741 ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac
     4801 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     4861 catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaatt cggttgaata
     4921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5041 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5101 aggcgtatca cgaccatggt tacaatttcc attcgccatt caggctgcgc aactgttggg
     5161 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     5221 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5281 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctctta cttgtacagc
     5341 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
     5401 cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc
     5461 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
     5521 ctgccgtcct cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg
     5581 tcggcggtga tatagacgtt gtggctgatg tagttgtact ccagcttgtg ccccaggatg
     5641 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     5701 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
     5761 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     5821 gggtagcggc tgaagcactg cacgccccag gtcagggtgg tcacgagggt gggccagggc
     5881 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
     5941 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     6001 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatgct taatttctcc
     6061 tctttagtta gttagcggcc gctctagaac tagtggatcc cccgggtaag taagtaatta
     6121 aagaggagaa attaagcatg cgtaaaggag aagaactttt cactggagtt gtcccaattc
     6181 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     6241 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     6301 ttccatggcc aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc
     6361 cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     6421 aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg
     6481 aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa
     6541 acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag
     6601 acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa
     6661 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     6721 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     6781 gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
     6841 aactatacaa ataagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga
     6901 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     6961 gtgagaatcc aagctagctt ggcgagattt tcagggagga attcgatatc aagcttatcg
     7021 ataccgtcga cctcgagggg gggccctaag taagtaaaaa gaggagaaat taagcatggt
     7081 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     7141 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     7201 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     7261 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     7321 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg aacgcgtgat
     7381 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     7441 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     7501 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     7561 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     7621 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     7681 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     7741 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtgagggt acccagcttt
     7801 tgttcccttt agtgagggtt aattgctggg cgctcttccg cttcctcgct cactgactcg
     7861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8221 tgtaggtatc tctgttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     8461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8761 acttgtgatc ccccctggag gatgccgtag ac
//
