LOCUS       40924_48263        4730 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pYS1R1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4730)
  AUTHORS   Shibayama Y, Dabbs ER, Yazawa K, Mikami Y.
  TITLE     Functional analysis of a small cryptic plasmid pYS1 from Nocardia
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4730)
  AUTHORS   Shibayama Y, Dabbs ER, Yazawa K, Mikami Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2010) School of Molecular and Cell Biology, 
            University of the Witwatersrand, 1 Jan Smuts Avenue, Johannesburg, 
            Gauteng 2050, South Africa
REFERENCE   3  (bases 1 to 4730)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4730)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-NOV-2010) School of Molecular and Cell Biology, University of 
            the Witwatersrand, 1 Jan Smuts Avenue, Johannesburg, Gauteng 2050, 
            South Africa"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(431..1474)
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiation protein"
                     /function="replication initiation in Gram-positive host"
                     /label=rep
                     /protein_id="ADR00353.1"
                     /translation="MKSCGWEPIANGIQIRLSATDGRMVAGFGGLETCGRIWLCPVCSA
                     KVRVRRGDQIAEACARHLDNEGAACWFITVTLPHEKGDALKDSFTALNEAWRYVKTGKA
                     YQADKKRFGILGDIRAVEVTHGENGWHPHAHILVFTTKQLEFDEMCAWAARLDARWARG
                     LAKVGWPQGKPGVRFTMVPVTKGKGVAAYVAKVQEKGLGNEIARADMKDATSGNRTPFG
                     ILADFGNDALADDLELWWEYEAATAGRSAIRWSPGLWAKLMPDEDELTDEEIAAEDEKG
                     STIAYMAPWTWRRLRTIQGAEIAVLEAVEAHGWGGLLRVLIDLRIGTQGVFTPDEWATP
                     GTVDLVG"
     gene            complement(431..1474)
                     /gene="rep"
                     /label=rep
                     /note="from pYS1"
     CDS             2434..3225
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      3512..4100
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4388..4409
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4424..4454
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4462..4478
                     /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)."
     primer_bind     4486..4502
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4512..4568
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4572..4588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgataagcta gccgatgggg ctgaccgtta ccgcggctgc gaaccggtcc gcgtcgaggc
       61 tggcccgaga gacctgggcg gaggtctggc cggatcggcc atagccgcgg ggagtcgtcg
      121 ggccggtcga gcattcggcc ggcctttttc gtgcgcgggg tggtggggtc gggcttcccc
      181 gggcctcgat tgtccagccc ttccgatgga gtcaagggac gcttcgctct tcgcccttga
      241 caccctctcc agtgctttgg atctggcctg tatcggggaa gggagtacgg gggtcaccga
      301 tggtggaaga ggtgtgtacc cgggcggcag ttccgagcgg ggaaggggga gcagcggagg
      361 gtcacacctc atgctccccg gtaacggtcc aatctccgct acacctcttg caccatcagt
      421 gaatccggag tcagccgacg agatccacag ttcccggggt ggcccattcg tcgggcgtga
      481 agactccctg agtaccgatc cggaggtcga tgagcacccg gagtaggccg ccccatccgt
      541 gggcctctac ggcctccagg acggcgattt cggctccctg gatagtccgg agccgccgcc
      601 acgtccacgg ggccatgtaa gcgattgtgg agcccttctc gtcctcggcc gcgatctcct
      661 cgtcggtcag ctcgtcctca tcgggcatga gcttggccca caggccgggg ctccagcgga
      721 tcgcggatcg gccggcggtt gcggcttcgt attcccacca gagttcgaga tcgtcggcca
      781 gggcgtcgtt tccgaagtcg gcgaggattc cgaacggggt gcggttcccg ctggtcgcgt
      841 ccttcatgtc cgctcgggcg atctcgttgc cgagtccctt ttcctgcacc ttggcgacgt
      901 aggccgctac gcccttgccc ttggtgaccg ggaccatcgt gaaccgaaca ccgggtttgc
      961 cctgcggcca gccgactttg gccagtcccc gagcccagcg ggcatcgagc cgggccgccc
     1021 acgcacacat ctcgtcgaat tcgagctgtt tcgtggtgaa caccaggatg tgggcgtgcg
     1081 ggtgccagcc gttctcgccg tgggtaacct cgacggcccg gatgtcgccg aggatcccga
     1141 accgcttctt gtcagcctgg taggccttgc cggtcttcac gtaccgccac gcctcattca
     1201 gcgcggtgaa cgagtccttg agcgcgtcgc ctttctcgtg cggcagcgtg accgtgatga
     1261 accagcaggc cgcgccctcg ttgtcgaggt gccgcgcgca agcctccgcg atctgatcac
     1321 cgcgtcgcac gcggactttg gcgctgcaca ccgggcagag ccagattcga ccgcaggtct
     1381 ccaggcctcc gaagcccgcg accatccggc cgtcggtggc gctcagcctg atctggattc
     1441 cgttggcgat tggttcccat ccgcagctct tcacccggtc gactgtcgag acctctcgga
     1501 gcaggtgtcg caggcggtat cggagtttcc gccgggccga aatggggtcc tgacctgcgg
     1561 gtgacgcaaa gtttgcgtag taccaagtcc ggaaactctc gccggggtgt tccaaggcct
     1621 ccccgcctcc gggcaaaccc gcctgcggcg ggcgcccgtc ggccaccgct cctgctaccg
     1681 cccgctcagc cgacggcgac cctccagcgg ggaggccaga gggggtgacc tgggattttg
     1741 tacgagagaa ggccttgcgg cacgcctcgt tgcagtaccg gcgaggtcgt ccggatttgg
     1801 ccgaccggag ttccgtgccg cacgcgacac agcttcggcg gtccgcaggc ccgttatcgt
     1861 gcgcagttgc taccatcgat cggtctcctt cagtgggagc acccatccgg cccgtgctgg
     1921 ctaggcaaga aggccggata agccccgagg gcgaaccagt gcattggttc gccctcagtg
     1981 cgtttcgggg tgctatgcgg gggtgaggct agttctcgtc gacgtccgac ggtgcgagca
     2041 acgccgcagg ggagtcgggc aatgtgccga gatgtgctag cttcacgctg ccgcaagcac
     2101 tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg
     2161 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca
     2221 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta
     2281 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
     2341 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
     2401 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2461 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2521 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2581 aagaccgacc tgtccggtgc cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg
     2641 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2701 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2761 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2821 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2881 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2941 ctgttcgcca ggctcaaggc gcggatgccc gacggcgagg atctcgtcgt gacccatggc
     3001 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3061 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3121 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3181 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3241 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3301 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3361 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caaaaggatc taggtgaaga
     3421 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3481 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3541 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3601 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3661 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3721 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3781 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3841 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3901 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3961 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4021 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4081 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4141 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4201 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4261 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4321 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     4381 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     4441 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     4501 accatgatta cgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat
     4561 gcaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4621 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4681 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
//
