LOCUS       Exported                9719 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses dcas9 (Streptococcus pasteurianus); repressed with 0.2% 
            glucose, induced with 0.3% xylose; for homologous recombination at 
            the xylose locus in Caulobacter crescentus; tetracycline resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pXGFPC-5 Pxyl-dcas9 (Spa)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9719)
  AUTHORS   Guzzo M, Castro LK, Reisch CR, Guo MS, Laub MT.
  TITLE     A CRISPR Interference System for Efficient and Rapid Gene Knockdown 
            in Caulobacter crescentus
  JOURNAL   mBio
REFERENCE   2  (bases 1 to 9719)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9719
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     66..82
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      83..129
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2656..2682
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6227..6243)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6254..6270)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(6468..7056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6548..6567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7162..7178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(7275..8474)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             8580..9230
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     oriT            9524..9633
                     /note="incP origin of transfer"
ORIGIN
        1 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaaaga tccgtcgacg
       61 gccgctctag aactagtgga tccaaataaa acgaaaggct cagtcgaaag actgggcctt
      121 tcgttttatc tgttgtttgt cgcattatac gcaaggcgac aaggtgctga tgccgctggc
      181 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
      241 acaacagttt ttatataagc ttagcgctac cggactcaga tccagatcga aatgtaccgg
      301 ctgaactggt ggaagcacgt tgagcctcgt caaggactca tgaagcgcgt ccctgctctc
      361 agtcgcctgc tcgttacgcc tgaggtgtca aaacaccgtc tgttcatttg gctagacgct
      421 agagttcttc cggatcacaa gctccaggtc gttacactcg atgacgactg ctcattcgga
      481 gttttacact caaggttcca cgaggtgtgg gctcttgcag cggggtcgtg gcacggctca
      541 ggcaatgatc cgcgctacac catctccacc accttcgaga ccttcccctt cccggaaggc
      601 ctgacgccca acatagccgc cgtcgactat gagggcgacc cgcgcgccca ggccatcgcc
      661 gcggcggcgg ccgagctgaa ccggctgcgg gaagcctggc tcaatccgcc cgacctggtg
      721 cggatcgagc ccgaggtcgt gcccggctat cccgaccgcg tgctgcccgt cagccccgag
      781 gcgggcgccg agctgaagaa gcgcaccctg accaacctct acaaccagcg cccggcctgg
      841 ctggacatgg cccaccagcg cctggacgcc gccgtcgccg ccgcctacgg ctggcccgac
      901 ggcctgaccg acgacgagat cctggagcgc ctgttcgccc tcaaccagga gcgcgcggcg
      961 gcggggcggt aggttagacc cctccaaaac cccgacctcc ccggcgaagg ccggggtcca
     1021 gttacatccg gaacgcttgg agtgggcccg cctacggcct cgaccaaccc gcagtcggca
     1081 aggggttcga tctggacccc ggcattcgcc ggggaggtcg gcagagaggt cagaaccgcg
     1141 tcttgcagcc ctgcttcaga tgaaccccgc gcaggaagcc gcggcgggcc aggccggcgt
     1201 cgacggcggc gttcatctgg cggtcggcgg gggcagcgcc ggtgacttcg cgcaccaggc
     1261 cccggaacgg gatgatgctg ttgatcaggg tcttgccacc ggcctcggcc agcttgccgg
     1321 ctcggttttc ggcgtactcg gcgccgacct tgtagtcggg ccccagcgcc gcgttcagcg
     1381 ccttcacctc attggccagg gcctggcagg acttcagacc cgcgaggctg taggggtcgt
     1441 caaaggcctt ggccaggacc ggcggcacct cgcgcttgga caggccgacg tcgcgcaccg
     1501 gctgggtgcc gatgtcgacg gccttcttgc ccgcctcgcc ccagcggccg gacggttcct
     1561 cgggcttggc cggcggcttg acctgggcgt gggcgctgga ggccagcagc acggcggcgg
     1621 ctccggccat gatcagcagc ttgttcggca tcaaaagcgc cctcccgaca aaacacgcca
     1681 cgggaacggc aagcctcgcc gccggttccc gcccgcaagt ccctcggcac gaactctaag
     1741 gcctgcgcac gatccatcgc caccacaaag cgtgtcatcc cggccgaagc gcagcggaga
     1801 gccgggaccc agggccacaa gcacggcgct tcgtcacccc ctgggtcccg gatagcgctc
     1861 cgcgcttccg ggatgacacg ggtttaagca cttgggtgag aagccctagt tcggcgcgcc
     1921 gcccgcttcc aggtcgaacg cggcgtcgaa ggcgtcgtcg ccaggaaccg tcccttccag
     1981 ggcggcgtcg aacaggtctt cgagcgatga ccagatgaag gccatctgct ccatgatcgt
     2041 cgggacgctg gccggatcga agccggggta gaggtcttcc accagccaca ggcccgtgcc
     2101 gggatcgaag gcgaagcggc agccgatcag gcggaactgg gcgcgggcca tggtctcgaa
     2161 cagggccgtc aggtcgcggg cggcgtccag gtcgtcgtgg tccagcacca ggcagcggat
     2221 ctgcagccag ccgtggtcgg gcagcaggta gaaggcgccc tcgtcctgat cctcgcccga
     2281 aacctccagc ccccggtcga tggcttcgac gacatagccg gccgcgcggc aggtgtcggt
     2341 gagcgcggcc agcagggcgg cttcctggtc aggggtcagg tcggtcatgg gcaagaggtc
     2401 caggtcgtgg tttgtcggcg gcttctagca tggaccgccc gcgcccgtga ggccgaggat
     2461 ttcgcgctgg tcagacaacc tacttgccgt ccccacatgt tagcgctacc aagtgccgac
     2521 gaacgcgcgc cgccgacggt gtcggcgctt cagacgctcg agttttgggg agacgaccat
     2581 atgcctgcag gcgccttaat taatatgcat ggtaccttaa gatctcgagc tcctgactca
     2641 ggaggtatcg atatgtaccc atacgatgtt ccagattacg ctatgaccaa cggcaagatc
     2701 ctgggcctgg ccatcggcat cgcctcggtg ggcgtgggca tcatcgaggc caagaccggc
     2761 aaggtggtgc acgccaactc gcgcctgttc tcggccgcca acgccgagaa caacgccgag
     2821 cgccgcggct tccgcggctc gcgccgcctg aaccgccgca agaagcaccg cgtgaagcgc
     2881 gtgcgcgacc tgttcgagaa gtacggcatc gtgaccgact tccgcaacct gaacctgaac
     2941 ccgtacgagc tgcgcgtgaa gggcctgacc gagcagctga agaacgagga gctgttcgcc
     3001 gccctgcgca ccatctcgaa gcgccgcggc atctcgtacc tggacgacgc cgaggacgac
     3061 tcgaccggct cgaccgacta cgccaagtcg atcgacgaga accgccgcct gctgaagaac
     3121 aagaccccgg gccagatcca gctggagcgc ctggagaagt acggccagct gcgcggcaac
     3181 ttcaccgtgt acgacgagaa cggcgaggcc caccgcctga tcaacgtgtt ctcgacctcg
     3241 gactacgaga aggaggcccg caagatcctg gagacccagg ccgactacaa caagaagatc
     3301 accgccgagt tcatcgacga ctacgtggag atcctgaccc agaagcgcaa gtactaccac
     3361 ggcccgggca acgagaagtc gcgcaccgac tacggccgct tccgcaccga cggcaccacc
     3421 ctggagaaca tcttcggcat cctgatcggc aagtgcaact tctacccgga cgagtaccgc
     3481 gcctcgaagg cctcgtacac cgcccaggag tacaacttcc tgaacgacct gaacaacctg
     3541 aaggtgtcga ccgagaccgg caagctgtcg accgagcaga aggagtcgct ggtggagttc
     3601 gccaagaaca ccgccaccct gggcccggcc aagctgctga aggagatcgc caagatcctg
     3661 gactgcaagg tggacgagat caagggctac cgcgaggacg acaagggcaa gccggacctg
     3721 cacaccttcg agccgtaccg caagctgaag ttcaacctgg agtcgatcaa catcgacgac
     3781 ctgtcgcgcg aggtgatcga caagctggcc gacatcctga ccctgaacac cgagcgcgag
     3841 ggcatcgagg acgccatcaa gcgcaacctg ccgaaccagt tcaccgagga gcagatctcg
     3901 gagatcatca aggtgcgcaa gtcgcagtcg accgccttca acaagggctg gcactcgttc
     3961 tcggccaagc tgatgaacga gctgatcccg gagctgtacg ccacctcgga cgagcagatg
     4021 accatcctga cccgcctgga gaagttcaag gtgaacaaga agtcgtcgaa gaacaccaag
     4081 accatcgacg agaaggaggt gaccgacgag atctacaacc cggtggtggc caagtcggtg
     4141 cgccagacca tcaagatcat caacgccgcc gtgaagaagt acggcgactt cgacaagatc
     4201 gtgatcgaga tgccgcgcga caagaacgcc gacgacgaga agaagttcat cgacaagcgc
     4261 aacaaggaga acaagaagga gaaggacgac gccctgaagc gcgccgccta cctgtacaac
     4321 tcgtcggaca agctgccgga cgaggtgttc cacggcaaca agcagctgga gaccaagatc
     4381 cgcctgtggt accagcaggg cgagcgctgc ctgtactcgg gcaagccgat ctcgatccag
     4441 gagctggtgc acaactcgaa caacttcgag atcgacgcca tcctgccgct gtcgctgtcg
     4501 ttcgacgact cgctggccaa caaggtgctg gtgtacgcct ggaccaacca ggagaagggc
     4561 cagaagaccc cgtaccaggt gatcgactcg atggacgccg cctggtcgtt ccgcgagatg
     4621 aaggactacg tgctgaagca gaagggcctg ggcaagaaga agcgcgacta cctgctgacc
     4681 accgagaaca tcgacaagat cgaggtgaag aagaagttca tcgagcgcaa cctggtggac
     4741 acccgctacg cctcgcgcgt ggtgctgaac tcgctgcagt cggccctgcg cgagctgggc
     4801 aaggacacca aggtgtcggt ggtgcgcggc cagttcacct cgcagctgcg ccgcaagtgg
     4861 aagatcgaca agtcgcgcga gacctaccac caccacgccg tggacgccct gatcatcgcc
     4921 gcctcgtcgc agctgaagct gtgggagaag caggacaacc cgatgttcgt ggactacggc
     4981 aagaaccagg tggtggacaa gcagaccggc gagatcctgt cggtgtcgga cgacgagtac
     5041 aaggagctgg tgttccagcc gccgtaccag ggcttcgtga acaccatctc gtcgaagggc
     5101 ttcgaggacg agatcctgtt ctcgtaccag gtggactcga agtacaaccg caaggtgtcg
     5161 gacgccacca tctactcgac ccgcaaggcc aagatcggca aggacaagaa ggaggagacc
     5221 tacgtgctgg gcaagatcaa ggacatctac tcgcagaacg gcttcgacac cttcatcaag
     5281 aagtacaaca aggacaagac ccagttcctg atgtaccaga aggactcgct gacctgggag
     5341 aacgtgatcg aggtgatcct gcgcgactac ccgaccacca agaagtcgga ggacggcaag
     5401 aacgacgtga agtgcaaccc gttcgaggag taccgccgcg agaacggcct gatctgcaag
     5461 tactcgaaga agggcaaggg caccccgatc aagtcgctga agtactacga caagaagctg
     5521 ggcaactgca tcgacatcac cccggaggag tcgcgcaaca aggtgatcct gcagtcgatc
     5581 aacccgtggc gcgccgacgt gtacttcaac ccggagaccc tgaagtacga gctgatgggc
     5641 ctgaagtact cggacctgtc gttcgagaag ggcaccggca actaccacat ctcgcaggag
     5701 aagtacgacg ccatcaagga gaaggagggc atcggcaaga agtcggagtt caagttcacc
     5761 ctgtaccgca acgacctgat cctgatcaag gacatcgcct cgggcgagca ggagatctac
     5821 cgcttcctgt cgcgcaccat gccgaacgtg aaccactacg tggagctgaa gccgtacgac
     5881 aaggagaagt tcgacaacgt gcaggagctg gtggaggccc tgggcgaggc cgacaaggtg
     5941 ggccgctgca tcaagggcct gaacaagccg aacatctcga tctacaaggt gcgcaccgac
     6001 gtgctgggca acaagtactt cgtgaagaag aagggcgaca agccgaagct ggacttcaag
     6061 aacaacaaga agtaaatact ctggccattt ttctttaccg cctttgagtg agctgatacc
     6121 gctcgccgca gccgaacgac cgagcgcaac gcgtgcggcc gctgattagc taagcagaag
     6181 gccatcctga cggatgcctt tttcgcttgc tagctgcagc ccgggggatc cactagttct
     6241 agagcggcca ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     6301 cccaacttaa tcgccttgca gcacatcccc ctttcgccag cggtatcagc tcactcaaag
     6361 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6421 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6661 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6901 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7021 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7081 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat acgaggccct
     7141 ttcgtcttca agaattaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     7201 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagca gatccgtccc
     7261 tatcgtttcc acgatcagcg atcggctcgt tgccctgcgc cgctccaaag cccgcgacgc
     7321 agcgccggca ggcagagcaa gtagagggca gcgcctgcaa tccatgccca cccgttccac
     7381 gttgttatag aagccgcata gatcgccgtg aagaggaggg gtccgacgat cgaggtcagg
     7441 ctggtgagcg ccgccagtga gccttgcagc tgcccctgac gttcctcatc cacctgcctg
     7501 gacaacattg cttgcagcgc cggcattccg atgccacccg aagcaagcag gaccatgatc
     7561 gggaacgcca tccatccccg tgtcgcgaag gcaagcagga tgtagcctgt gccgtcggca
     7621 atcattccga gcatgagtgc ccgcctttcg ccgagccggg cggctacagg gccggtgatc
     7681 attgcctggg cgagtgaatg cagaatgcca aatgcggcaa gcgaaatgcc gatcgtggtc
     7741 gcgtcccagt gaaagcgatc ctcgccgaaa atgacccaaa gcgcggccgg cacctgtccg
     7801 acaagttgca tgatgaagaa gaccgccatc agggcggcga cgacggtcat gccccgggcc
     7861 caccggaacg aagcgagcgg gttgagagcc tcccggcgta acggccggcg ttcgcctttg
     7921 tgcgactccg gcaaaaggaa acagcccgtc aggaaattga ggccgttcaa ggctgccgcg
     7981 gcgaagaacg gagcgtgggg ggagaaaccg cccatcagcc caccgagcac aggtcccgcg
     8041 accatcccga acccgaaaca ggcgctcatg aagccgaagt gccgcgcgcg ctcatcgcca
     8101 tcagtgatat cggcaatata agcgccggct accgccccag tcgccccggt gatgccggcc
     8161 acgatccgcc cgatatagag aacccaaagg aaaggcgctg tcgccatgat ggcgtagtcg
     8221 acagtggcgc cggccagcga gacgagcaag attggccgcc gcccgaaacg atccgacagc
     8281 gcgcccagca caggtgcgca ggcaaattgc accaacgcat acagcgccag cagaatgcca
     8341 tagtgggcgg tgacgtcgtt cgagtgaacc agatcgcgca ggaggcccgg cagcaccggc
     8401 ataatcaggc cgatgccgac agcgtcgagc gcgacagtgc tcagaattac gatcaggggt
     8461 atgttgggtt tcacgtctgg cctccggacc agcctccgct ggtccgattg aacgcgcgga
     8521 ttctttatca ctgataagtt ggtggacata ttatgtttat cagtgataaa gtgtcaagca
     8581 tgacaaagtt gcagccgaat acagtgatcc gtgccgccct ggacctgttg aacgaggtcg
     8641 gcgtagacgg tctgacgaca cgcaaactgg cggaacggtt gggggttcag cagccggcgc
     8701 tttactggca cttcaggaac aagcgggcgc tgctcgacgc actggccgaa gccatgctgg
     8761 cggagaatca tacgcattcg gtgccgagag ccgacgacga ctggcgctca tttctgatcg
     8821 ggaatgcccg cagcttcagg caggcgctgc tcgcctaccg cgatggcgcg cgcatccatg
     8881 ccggcacgcg accgggcgca ccgcagatgg aaacggccga cgcgcagctt cgcttcctct
     8941 gcgaggcggg tttttcggcc ggggacgccg tcaatgcgct gatgacaatc agctacttca
     9001 ctgttggggc cgtgcttgag gagcaggccg gcgacagcga tgccggcgag cgcggcggca
     9061 ccgttgaaca ggctccgctc tcgccgctgt tgcgggccgc gatagacgcc ttcgacgaag
     9121 ccggtccgga cgcagcgttc gagcagggac tcgcggtgat tgtcgatgga ttggcgaaaa
     9181 ggaggctcgt tgtcaggaac gttgaaggac cgagaaaggg tgacgattga tcaggaccgc
     9241 tgccggagcg caacccactc actacagcag agccatgtag acaacatccc ctcccccttt
     9301 ccaccgcgtc agacgcccgt agcagcccgc tacgggcttt ttcatgccct gccctagcgt
     9361 ccaagcctca cggccgcgct cggcctctct ggcggccttc tggcgctcct gctgcggcgt
     9421 ccgctcgtat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     9481 aacgtggctt tccccccccc cccccaccgc ggtggcggcc gccccggcca gcctcgcaga
     9541 gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg
     9601 ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga
     9661 aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatac
//
