LOCUS       40924_4719        7277 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pARAtrfA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  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  JOURNAL   Unpublished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 7277)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-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 7277)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7277)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-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..7277
                     /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..5180
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             5192..5212
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5222..6367
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     protein_bind    complement(6387..6411)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6655..6741
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6833..6860
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        6879..6970
                     /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 aaagcaggct ccgcggccgc cttgtttaac tttaagaagg agcccttcac
     5221 catgaatcgg acgtttgacc ggaaggcata caggcaagaa ctgatcgacg cggggttttc
     5281 cgccgaggat gccgaaacca tcgcaagccg caccgtcatg cgtgcgcccc gcgaaacctt
     5341 ccagtccgtc ggctcgatgg tccagcaagc tacggccaag atcgagcgcg acagcgtgca
     5401 actggctccc cctgccctgc ccgcgccatc ggccgccgtg gagcgttcgc gtcgtctcga
     5461 acaggaggcg gcaggtttgg cgaagtcgat gaccatcgac acgcgaggaa ctatgacgac
     5521 caagaagcga aaaaccgccg gcgaggacct ggcaaaacag gtcagcgagg ccaagcaggc
     5581 cgcgttgctg aaacacacga agcagcagat caaggaaatg cagctttcct tgttcgatat
     5641 tgcgccgtgg ccggacacga tgcgagcgat gccaaacgac acggcccgct ctgccctgtt
     5701 caccacgcgc aacaagaaaa tcccgcgcga ggcgctgcaa aacaaggtca ttttccacgt
     5761 caacaaggac gtgaagatca cctacaccgg cgtcgagctg cgggccgacg atgacgaact
     5821 ggtgtggcag caggtgttgg agtacgcgaa gcgcacccct atcggcgagc cgatcacctt
     5881 cacgttctac gagctttgcc aggacctggg ctggtcgatc aatggccggt attacacgaa
     5941 ggccgaggaa tgcctgtcgc gcctacaggc gacggcgatg ggcttcacgt ccgaccgcgt
     6001 tgggcacctg gaatcggtgt cgctgctgca ccgcttccgc gtcctggacc gtggcaagaa
     6061 aacgtcccgt tgccaggtcc tgatcgacga ggaaatcgtc gtgctgtttg ctggcgacca
     6121 ctacacgaaa ttcatatggg agaagtaccg caagctgtcg ccgacggccc gacggatgtt
     6181 cgactatttc agctcgcacc gggagccgta cccgctcaag ctggaaacct tccgcctcat
     6241 gtgcggatcg gattccaccc gcgtgaagaa gtggcgcgag caggtcggcg aagcctgcga
     6301 agagttgcga ggcagcggcc tggtggaaca cgcctgggtc aatgatgacc tggtgcattg
     6361 caaacgctag aagggtgggc gcgccgaccc agctttcttg tacaaagtgg tgatggggat
     6421 cctctagagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt
     6481 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     6541 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     6601 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     6661 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     6721 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     6781 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     6841 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     6901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7201 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7261 gaatgacttg gttgagt
//
