LOCUS       62056_18830        6376 bp DNA     circular SYN 28-FEB-2021
DEFINITION  Cloning vector pRGC, complete sequence.
ACCESSION   MW148406
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  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  JOURNAL   Unpublished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 6376)
  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 6376)
  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            "Unpublished"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..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(144..974)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      988..1339
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             complement(1745..2275)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(2464..2492)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(2510..2614)
                     /label=AmpR promoter
     primer_bind     2853..2869
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2879..2897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2915..3631)
                     /label=ECFP
                     /note="enhanced CFP"
     regulatory      complement(3638..3649)
                     /label=strong bacterial ribosome binding site
                     /note="strong bacterial ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
     primer_bind     3667..3683
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3704..3715
                     /label=strong bacterial ribosome binding site
                     /note="strong bacterial ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
     CDS             3722..4435
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     terminator      4458..4552
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4603..4619)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4642..4653
                     /label=strong bacterial ribosome binding site
                     /note="strong bacterial ribosome binding site"
                     /regulatory_class="ribosome_binding_site"
     CDS             4660..5367
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     promoter        complement(5390..5408)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      complement(5600..6188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcatgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
       61 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa aggcaggccg
      121 ggccgtggtg gccacggcct ctaggccaga tccagcggca tctgggttag tcgagcgcgg
      181 gccgcttccc atgtctcacc agggcgagcc tgtttcgcga tctcagcatc tgaaatcttc
      241 ccggccttgc gcttcgctgg ggccttaccc accgccttgg cgggcttctt cggtccaaaa
      301 ctgaacaaca gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc cacctgtagc
      361 gggctgtgct cgttgatctg cgtcacggct ggatcaagca ctcgcaactt gaagtccttg
      421 atcgagggat accggccttc cagttgaaac cactttcgca gctggtcaat ttctatttcg
      481 cgctggccga tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc gtgggtgctg
      541 tccatcttgg ccacgtcagc caaggcgtat ttggtgaact gtttggtgag ttccgtcagg
      601 tacggcagca tgtctttggt gaacctgagt tctacacggc cctcaccctc ccggtagatg
      661 attgtttgca cccagccggt aatcatcaca ctcggtcttt tccccttgcc attgggctct
      721 tgggttaacc ggacttcccg ccgtttcagg cgcagggccg cttctttgag ctggttgtag
      781 gaagattcga tagggacacc cgccatcgtc gctatgtcct ccgccgtcac tgaatacatc
      841 acttcatcgg tgacaggctc gctcctcttc acctggctaa tacaggccag aacgatccgc
      901 tgttcctgaa cactgaggcg atacgcggcc tcgaccaggg cattgctttt gtaaaccatt
      961 gggggtgagg ccacgttcga cattccttgt gtataagggg acactgtatc tgcgtcccac
     1021 aatacaacaa atccgtccct ttacaacaac aaatccgtcc cttcttaaca acaaatccgt
     1081 cccttaatgg caacaaatcc gtcccttttt aaactctaca ggccacggat tacgtggcct
     1141 gtagacgtcc taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac gcaaaaaacg
     1201 caccactacg tggccccgtt ggggccgcat ttgtgcccct gaaggggcgg gggaggcgtc
     1261 tgggcaatcc ccgttttacc agtcccctat cgccgcctga gagggcgcag gaagcgagta
     1321 atcagggtat cgaggcggat tcacccttgg cgtccaacca gcggcaccag cggcgcctga
     1381 gaggcctaca gagcggttgg acaccaaggg gaggggctaa gaccggttta tcagtcccct
     1441 ttccctcgtt tctttccaac gcgatagccc agcaaggccg ccaccgttgc caccgtcacc
     1501 ccagcaagca cagccagtgg cgtgtaattg gggtctgacg ctcagtggaa cgaaaactca
     1561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccatgcatca
     1621 gcagcaacag cagcaggggc tgcatacttg gtctgacaat cgatgcgaat tggccgcggc
     1681 gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt
     1741 gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact
     1801 ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag
     1861 caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc
     1921 tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa
     1981 acttgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa
     2041 gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt
     2101 gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg
     2161 atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta
     2221 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg
     2281 ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga
     2341 ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     2401 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
     2461 tacagtttac gaaccgaaca ggcttatgtc aattcggttg aatactcata ctcttccttt
     2521 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2581 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2641 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgacca
     2701 tggttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2761 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2821 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     2881 atacgactca ctatagggcg aattggagct cttacttgta cagctcgtcc atgccgagag
     2941 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     3001 ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     3061 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     3121 tgtggcggat cttgaagttg gccttgatgc cgttcttctg cttgtcggcg gtgatataga
     3181 cgttgtggct gatgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     3241 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     3301 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     3361 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     3421 actgcacgcc ccaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     3481 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     3541 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     3601 cggtgaacag ctcctcgccc ttgctcacca tgcttaattt ctcctcttta gttagttagc
     3661 ggccgctcta gaactagtgg atcccccggg taagtaagta attaaagagg agaaattaag
     3721 catgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     3781 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     3841 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     3901 tgtcactact ttcggttatg gtgttcaatg ctttgcgaga tacccagatc atatgaaaca
     3961 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
     4021 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     4081 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     4141 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     4201 aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga
     4261 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     4321 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     4381 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataagc
     4441 ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag aactccatct
     4501 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta
     4561 gcttggcgag attttcaggg aggaattcga tatcaagctt atcgataccg tcgacctcga
     4621 gggggggccc taagtaagta aaaagaggag aaattaagca tggtgagcaa gggcgaggag
     4681 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     4741 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4801 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     4861 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     4921 ttgaagctgt ccttccccga gggcttcaag tgggaacgcg tgatgaactt cgaggacggc
     4981 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     5041 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     5101 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     5161 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     5221 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     5281 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     5341 accggcggca tggacgagct gtacaagtga gggtacccag cttttgttcc ctttagtgag
     5401 ggttaattgc tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5461 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5521 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5581 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5641 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5701 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5761 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctctgtt
     5821 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5941 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6001 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6061 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6301 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgt gatcccccct
     6361 ggaggatgcc gtagac
//
