LOCUS       40924_4704        7726 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pARAGW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7726)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     An improved method for the oriT-directed cloning and 
            functionalization of large bacterial genomic regions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7726)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-AUG-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   3  (bases 1 to 7726)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7726)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-AUG-2012) Microbiology Immunology and Pathology, Colorado State 
            University, IDRC at Foothills Campus, Campus Delivery 0922, Fort 
            Collins, CO 80523-0922, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(44..874)
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYCLSVQEQRIVLASISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMGSTHAIRLYELLTQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKARKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      888..1239
                     /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)"
     promoter        1359..1406
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             1425..1796
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      2080..2535
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2646..3234
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3934..4809)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4836..5120
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     protein_bind    5156..5280
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5305..5335
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             5389..6045
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             6390..6692
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(6736..6860)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      7104..7190
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      7282..7309
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        7328..7419
                     /label=AmpR promoter
ORIGIN
        1 ctagtaacgg ccgccagtgt gctggaattc ggcttggcct ctaggccaga tccagcggca
       61 tctgggttag tcgagcgcgg gccgcttccc atgtctcacc agggcgagcc tgtttcgcga
      121 tctcagcatc tgaaatcttc ctggccttgc gcttcgcagg ggccttaccc accgccttgg
      181 cgggcttctt cggtccaaaa ctgaacaaca gatgtgtgac cttgcgcccg gtctttcgct
      241 gcgcccactc cacctgtagc gggctgtgct cgttgatctg cgtcacggct ggatcaagca
      301 ctcgcaactt gaagtccttg atcgagggat accggccttc cagttgaaac cactttcgca
      361 gctggtcaat ttctatttca cgctggccga tgctgtccca ttgcgtgagc agctcgtaaa
      421 gcctgatcgc gtgggtgctg cccatcttgg ccacgtcagc caaggcgtat ttggtgaact
      481 gtttggtgag ttccgtcaga tacggcagca tgtctttggt gaacctgagt tctacacggc
      541 cctcaccctc ccggtagatg attgtttgca cccagccggt aatcatcaca ctcggtcttt
      601 tccccttgcc attgggctct tgggttaacc ggacttcccg ccgtttcagg cgcagggccg
      661 cttctttgag ctggttgtag gaagattcga tagggacacc cgccatcgtc gctatgtcct
      721 ccgccgtcac tgaatacatc acttcatcgg tgacaggctc gctcctcttc acctggctaa
      781 tactggccag aacgatccgc tgttcctgaa cactgaggca atacgcggcc tcgaccaggg
      841 cattgctttt gtaaaccatt gggggtgagg ccacgttcga cattccttgt gtataagggg
      901 acactgtatc agcgtcccac aatacaacaa atccgtccct ttacaacaac aaatccgtcc
      961 cttcttaaca acaaatccgt cccttaatgg caacaaatcc gtcccttttt aaactctaca
     1021 ggccacggat tacgtggcct gtagacgtcc taaaaggttt aaaagggaaa aggaagaaaa
     1081 gggtggaaac gcaaaaaacg caccactacg tggccccgtt ggggccgcat ttgtgcccct
     1141 gaaggggcgg gggaggcgtc tgggcaatcc ccgttttacc agtcccctat cgccgcctga
     1201 gagggcgcag gaagcgagta atcagggtat cgaggcggat tcacccttgg cgtccaacca
     1261 gcggcaccag cggcgaattc acgaaaagcc gaattctgca gataattcgg cttcgacgcg
     1321 taactcacgt taagggattt tggtcatgat cagcacgtgt tgacaattaa tcatcggcat
     1381 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
     1441 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     1501 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     1561 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     1621 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     1681 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     1741 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgat
     1801 catgatgata ttattttatc ttgtgcaatg taacatcaga gataagccga attatccatc
     1861 acactggcgg ccgctcgagc atgcatctag tgggtctcgc ggtatcattg cagcactggg
     1921 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1981 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2041 gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat
     2101 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2161 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2221 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2281 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2341 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaacttgaa
     2401 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2461 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2521 taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2581 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2641 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2701 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2761 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2821 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2881 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2941 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3001 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3061 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3121 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3181 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3241 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3301 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3361 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3421 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agggtcatgg
     3481 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3541 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3601 cgtcatcacc gaaacgcgcg aggcagcaag gagatggcgc ccaacagtcc cccggccacg
     3661 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     3721 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     3781 atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcatcga
     3841 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
     3901 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
     3961 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     4021 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     4081 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     4141 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
     4201 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     4261 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     4321 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     4381 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     4441 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     4501 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     4561 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     4621 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     4681 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     4741 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     4801 ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc catattgcat
     4861 cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac cggtaacccc
     4921 gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa
     4981 aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg
     5041 ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca
     5101 actctctact gtttctccat acccgttttt ttgggctagc aggagggtac ccatcacaag
     5161 tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata tattaaatta
     5221 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg
     5281 gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg
     5341 attttgagtt aggatccggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     5401 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     5461 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     5521 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     5581 cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata
     5641 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     5701 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     5761 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     5821 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     5881 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     5941 atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg
     6001 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaaac gcgtggatcc
     6061 ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga
     6121 atatatactg atatgtatac ccgaagtatg tcaaaaagag gtgtgctatg aagcagcgta
     6181 ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat
     6241 ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg
     6301 gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc
     6361 ttttgctgac gagaacaggg actggtgaaa tgcagtttaa ggtttacacc tataaaagag
     6421 agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac
     6481 ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt
     6541 acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg
     6601 tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca
     6661 aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccgtt atacacagcc
     6721 agtctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt
     6781 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
     6841 gctttcttgt acaaagtggt gatggggatc ctctagagtc gacctgcagg catgcaagct
     6901 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     6961 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     7021 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     7081 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     7141 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     7201 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     7261 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     7321 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7381 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7441 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7501 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7561 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7621 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     7681 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagt
//
