LOCUS       pCarCBAE                7045 bp ds-DNA     circular SYN 17-SEP-2018
DEFINITION  synthetic circular DNA.
ACCESSION   . (modified)
VERSION     .
KEYWORDS    pHS-CarCBAE.
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
            .
REFERENCE   1  (bases 1 to 7045)
  AUTHORS   TUDelft
  TITLE     Direct Submission
  JOURNAL   Exported 7 Dec 2015 from SnapGene 2.8.3 http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7045
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             complement(49..1131)
                     /codon_start=1
                     /note="lacI"
                     /note="lacI does not contain internal EcoRV site"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
     promoter        1500..1575
                     /note="lacUV5 promoter"
     misc_binding    1516..1579
                     /note="lac operator"
     primer_bind     1529..1571
     primer_bind     1536..1555
     misc_feature    1555..1594
                     /note="Overlap(1)"
     misc_feature    complement(1559..1579)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1577..1600)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    1580..1594
                     /note="BglBrick 1(1)"
     primer_bind     1594..1614
     RBS             1595..1606
                     /note="Strong RBS"
     misc_feature    1607..2428
                     /note="CarC"
                     /note="Geneious type: CDS"
     RBS             2429..2440
                     /note="Strong RBS"
     misc_feature    2440..3192
                     /note="CarB"
                     /note="Geneious type: CDS"
     misc_feature    3193
                     /note="BglBrick 2(1)"
     primer_bind     3194..3216
     misc_feature    3194..3198
                     /note="BglBrick 1"
     misc_feature    3194..3198
                     /note="Overlap"
     RBS             3199..3210
                     /note="Strong RBS"
     CDS             3211..4722
                     /codon_start=1
                     /note="CarA"
                     /translation="MSNSFCVVYKGSDTDINNIQRDFDGKGEALSNGYLFIEQNGHYQ
                     KCEMERGTAYLIGSLYNRTFLIGLAGVWEGEAYLANDAELLALLFTRLGANALALAEG
                     DFCFFIDEPNGELTVITESRGFSPVHVVQGKKAWMTNSLKLVTAAEGEGALWFEEEAL
                     VCQSLMRADTYTPVKNAQRLKPGAVHVLTHDSEGYSFVESRTLTTPASNQLLALPREP
                     LLALIDRYLNAPLEDLAPRFDTVGIPLSGGLDSSLVTALASRHFKKLNTYSIGTELSN
                     EFEFSQQVADALGTHHQMKILSETEVINGIIESIYYNEIFDGLSAEIQSGLFNVYRQA
                     QGQVSCMLTGYGSDLLFGGILKPGAQYDNPNQLLAEQVYRTRWTGEFATHGASCYGID
                     IRHPFWSHSLISLCHALHPDYKIFDNEVKNILREYADSLQLLPKDIVWRKKIGIHEGS
                     SVNQAFANVLGSTVDNYQTKSRFTYRVYQAFLRGRLSITDVTPSQLKDLIKKD"
     primer_bind     4692..4751
     misc_feature    complement(4707..4728)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    4729..4744
                     /note="carD (involved in carbapenem biosynthesis) AAD3"
     CDS             4744..5022
                     /codon_start=1
                     /note="carE (Probable ferredoxin, 2F3-2S) AAD38233.1"
                     /translation="MSKFHCHIRDTAIGFPVSEDESVLSSAYEAGVELPYRCASGYCG
                     VCKVRLTSGNVNMDHSGGISRKDIADGYILPCCSVPLSNLEIEPVSSC"
     primer_bind     4971..4992
     primer_bind     4985..5044
     misc_feature    complement(5002..5022)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    5016..5022
                     /note="carF (involved in resistance to carbapenem) AAD"
     primer_bind     5023..5043
     misc_feature    5023..5024
                     /note="BglBrick 2(1)"
     misc_feature    complement(5028..5049)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(5030..5051)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(5032..5051)
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     5034..5054
     misc_feature    5034..5162
                     /note="dbl term"
     misc_feature    complement(5038..5059)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(5054..5073)
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(5148..5167)
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     5189..5208
     misc_feature    complement(5195..5214)
                     /note="Geneious type: primer_bind_reverse"
     rep_origin      complement(5296..5978)
     terminator      5984..6089
                     /note="TO"
     misc_feature    complement(6115..6909)
                     /note="Kan/neoR"
ORIGIN      
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg
     1561 tgagcggata acaatttcag aattcaaaag atctaggagg aaaaaaatgt ccgagatcgt
     1621 gaaatttaac cctgtgatgg cttctggctt cggcgcatac attgatcacc gtgacttcct
     1681 ggaagctaaa acggaaacga tcaaaaacct gctgatgcgc caaggtttcg tggttgtcaa
     1741 aaacctggat attgactccg atactttccg tgatatctac tctgcatacg gtaccatcgt
     1801 cgaatacgcg gatgaaaaaa ttggtgttgg ttttggttat cgcgataccc tgaaactgga
     1861 aggcgaaaaa ggtaaaatcg ttaccggtcg tggtcaactg cctttccacg ctgacggtgg
     1921 tctgctgctg tctcaggtcg atcaggtttt cctgtacgcg gcggagatta aaaacgtcaa
     1981 attccgcggt gcaactaccg tttgcgatca cgctctggcg tgccaggaaa tgccagcaca
     2041 cctgctgcgc gttctggaag aggagacgtt cgaagttcgt gtgctggaac gtggttacta
     2101 cgtggatgtg tcccctgatg gttggttcaa agtgccggtg ttcactgatc tgggttgggt
     2161 tcgtaaaatg ctgatctatt tcccattcga tgaaggccag ccggcaagct gggaaccgcg
     2221 tattgttggt tttaccgacc acgaaaccca ggcattcttt caagaactgg gtgcgttcct
     2281 gaaacagccg cgttattact acaaacattt ctgggaagac ggtgacctgc tgattatgga
     2341 taaccgtcgt gttatccacg agcgtgaaga atttaacgat gatgacatcg ttcgtcgtct
     2401 gtaccgcggc caaactgcag atatctaaag gaggaaaaaa tggtattcga agaaaactcc
     2461 gatgaagtac gcgtgattac cctggatcac ccgaacaaac ataacccatt ttcccgtacc
     2521 ctggaaacta gcgtgaagga cgctctggct cgtgcaaacg cggatgatag cgtgcgtgcg
     2581 gtcgttgtgt atggcggcgc tgaacgtagc ttcagcgcag gcggtgattt caacgaagtc
     2641 aagcagctgt ctcgctccga agatattgaa gaatggatcg atcgtgtgat cgacctgtat
     2701 caggcagtcc tgaacgtaaa caaaccgacg atcgcggccg ttgatggcta cgcgatcggt
     2761 atgggcttcc aattcgctct gatgtttgac cagcgcctga tggccagcac cgcgaacttc
     2821 gtgatgccgg aactgaaaca cggtatcggt tgctctgtag gtgcagcaat cctgggtttc
     2881 acccatggtt tctccaccat gcaagaaatc atctaccagt gccagtctct ggatgcaccg
     2941 cgctgcgtcg actaccgcct ggtgaatcag gtcgtagagt cttccgcgct gctggacgcg
     3001 gcgatcactc aggcgcacgt gatggcgtcc tacccggctt ccgcattcat taacacgaag
     3061 cgtgcggtaa acaagccgtt cattcacctg ctggaacaga cccgtgatgc ttccaaagca
     3121 gtgcacaaag ccgcattcca ggcgcgtgat gcccagggtc acttcaaaaa cgttctgggt
     3181 aagaaatact gaggatctag gaggaaaaaa atgagcaata gcttctgcgt cgtttacaaa
     3241 ggttccgata ctgacattaa caacattcag cgtgatttcg acggtaaagg cgaagccctg
     3301 tctaacggct atctgttcat cgaacagaac ggccactacc agaaatgcga aatggagcgt
     3361 ggcacggcgt acctgattgg ttctctgtat aaccgtactt tcctgattgg cctggcgggc
     3421 gtttgggaag gtgaagcata cctggctaac gatgctgaac tgctggccct gctgtttacg
     3481 cgcctgggtg ccaacgcact ggctctggcg gaaggcgatt tctgcttttt cattgatgaa
     3541 ccgaatggtg agctgaccgt tatcacggaa tctcgcggct tctccccggt ccatgttgtt
     3601 caaggtaaga aggcatggat gacgaacagc ctgaaactgg taactgctgc cgaaggcgag
     3661 ggcgcactgt ggtttgaaga agaagctctg gtgtgccagt ctctgatgcg tgccgatact
     3721 tatacgccgg ttaagaacgc gcagcgtctg aaaccgggcg cagtgcacgt tctgacccac
     3781 gatagcgaag gctacagctt tgtagagtct cgtaccctga ccactccggc atccaaccaa
     3841 ctgctggctc tgccgcgtga accgctgctg gcactgatcg atcgctacct gaacgctccg
     3901 ctggaggacc tggcaccacg cttcgacacg gttggtatcc cgctgagcgg tggtctggac
     3961 tcttccctgg tgactgccct ggctagccgc catttcaaga aactgaacac gtattccatc
     4021 ggtactgagc tgtccaacga gtttgagttc agccagcagg ttgcagatgc tctgggtacg
     4081 caccaccaga tgaaaatcct gtccgaaacg gaagttatta acggtatcat cgagtccatt
     4141 tactacaacg aaatcttcga cggtctgagc gcggaaatcc agagcggtct gttcaacgtg
     4201 taccgtcagg ctcagggcca agtctcctgc atgctgactg gctatggttc tgacctgctg
     4261 ttcggcggca tcctgaaacc tggcgctcag tatgataatc cgaaccagct gctggcggaa
     4321 caagtgtacc gcacccgttg gaccggcgaa tttgcgaccc acggtgcgtc ttgctacggc
     4381 atcgatatcc gccacccgtt ttggtcccac agcctgatta gcctgtgcca cgccctgcac
     4441 cctgattaca agattttcga taatgaagtt aaaaacatcc tgcgtgaata cgctgatagc
     4501 ctgcagctgc tgccaaagga catcgtttgg cgtaaaaaga tcggtattca cgaaggttct
     4561 tctgttaatc aggcgttcgc aaacgtactg ggttctacgg ttgacaatta ccagaccaaa
     4621 tctcgtttca cttatcgtgt ttaccaggcg ttcctgcgcg gtcgcctgtc catcactgac
     4681 gttaccccgt cccagctgaa ggatctgatc aaaaaggact gaggatcccg taggagggat
     4741 gtaatgtcaa aatttcattg tcatattcgc gatacggcta tcggtttccc cgtgtcggag
     4801 gatgaaagcg tactcagttc agcctatgaa gctggtgtcg aactgcctta tcgctgtgct
     4861 tctggttact gtggtgtatg caaggtgcgc ctgacgtccg gcaacgtcaa catggatcat
     4921 tctggtggga tctcacgtaa agatatcgct gatggttaca ttctaccttg ctgttctgtg
     4981 ccgctctcta acctcgaaat tgaacctgta tcgtcatgct gaagtaagga tctccaggca
     5041 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     5101 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     5161 tacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5941 gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa
     6001 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     6061 ttactggatc tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa
     6121 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     6181 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     6241 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     6301 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     6361 atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc
     6421 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     6481 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     6541 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     6601 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     6661 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     6721 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     6781 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     6841 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     6901 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca
     6961 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     7021 agagggcgcc ccagctggca attcc
//
