LOCUS       sequence-74538-e        4963 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4963)
  AUTHORS   Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, 
            Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA
  TITLE     Design of orthogonal genetic switches based on a crosstalk map of 
            sigmas, anti-sigmas, and promoters.
  JOURNAL   Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.
  PUBMED    24169405
REFERENCE   2  (bases 1 to 4963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2013-10-29"; volume: "9:702. doi"; pages: " 
            10.1038/msb.2013.58"
COMMENT     Sequence Label: pVRc03_1244
FEATURES             Location/Qualifiers
     source          1..4963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(448..465)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      618..704
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      796..823
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        842..933
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1103..1762
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1169..1188)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1904..2359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1991..2010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3045..3062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3179..3724)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3259..3278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3796..3815
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4946..4957
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaaacagt tcgatccgga tgttaacatt cgtccgaaag aagatagcat tcgtcattat
       61 agcgttccgg atgattattt tgcaagcttc accgataaac tgatggcaca gattccggca
      121 gttaaagaag aaaataccgt tgttgttccg agcgttgcac tgtggccgaa actgcgtccg
      181 ctgctgtatc tggcagcaag ctttctgctg atgattggta tgtttaaagc ctttagcctg
      241 tttggtgttg gtagcgatac cggtcgtacc accagtacca gcgcaggtct ggttgcactg
      301 gatcatggtg atacccgttg gagcgaagat accgattatc gtgattttct gcatgataat
      361 tgtgccgaaa ccgttagtga tgaatgggtt ctgaccgatt ttagcgaata aggatccctg
      421 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
      481 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
      541 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
      601 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      661 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
      721 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
      781 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
      841 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      901 taaatgcttc aataatattg aaaaaggaag actatgagta ttcaacattt ccgtgtctgc
      961 ttacataaac agtaatacaa ggggtgttta ctagaggttg atcgggcacg taagaggttc
     1021 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
     1081 ttcaggagct aaggaagcta aaatggagaa aaaaatcacg ggatatacca ccgttgatat
     1141 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1201 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1261 caagttttat ccggccttta ttcacattct tgcccgcctg atgaacgctc acccggagtt
     1321 tcgtatggcc atgaaagacg gtgagctggt gatctgggat agtgttcacc cttgttacac
     1381 cgttttccat gagcaaactg aaacgttttc gtccctctgg agtgaatacc acgacgattt
     1441 ccggcagttt ctccacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1501 tttccctaaa gggtttattg agaatatgtt ttttgtctca gccaatccct gggtgagttt
     1561 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcacgat
     1621 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgatcc aggttcatca
     1681 tgccgtttgt gatggcttcc atgtcggccg catgcttaat gaattacaac agtactgtga
     1741 tgagtggcag ggcggggcgt aataatacta gctccggcaa aaaaacgggc aaggtgtcac
     1801 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgtttgccac ctgacgtcta
     1861 agctgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     1921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1981 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2041 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2101 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2221 tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     2281 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2341 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2401 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2461 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2521 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2581 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2641 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2701 agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     2761 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     2821 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg
     2881 cgtttaaggg caccaataac tgccttaaaa aaagctagcg gagtgtatac tggcttacta
     2941 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     3001 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     3061 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     3121 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     3181 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     3241 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     3301 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     3361 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     3421 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3481 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     3541 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     3601 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     3661 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     3721 tcaagaagat catcttatta atcagataaa atatttgctc atgagcccga agtggcgagc
     3781 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     3841 ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatca
     3901 tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
     3961 ccctcgagcc gtcaattgtc tgattcgtta ccaattatta atttttaaag tatgggcaat
     4021 caattgctcc tgttaaaatt gctttagaaa tactttggca gcggtttgtt gtattgagtt
     4081 tcatttgcgc attggttaaa tggaaagtga cagtacgctc actgcaacct aatatttttg
     4141 aaatatccca agagcttttt ccttcgcatg cccacgctaa acattctttt tctcttttgg
     4201 ttaaatcgtt gtttgattta ttatttgcta tatttatttt tcgataatta tcaactagag
     4261 aaggaacaat taatggtatg ttcatacacg catgtaaaaa taaactatct atatagttgt
     4321 ctttttctga atgtgcaaaa ctaagcattc cgaagccatt gttagccgta tgaataggga
     4381 aactaaaccc agtgataaga cctgatgttt tcgcttcttt aattacattt ggagattttt
     4441 tatttacagc attgttttca aatatattcc aattaattgg tgaatgattg gagttagaat
     4501 aatctactat aggatcatat tttattaaat tagcgtcatc ataatattgc ctccattttt
     4561 tagggtaatt atctaggatt gaaatatcag atttaaccat agaatgagga taaatgatcg
     4621 cgagtaaata atattcacaa tgtaccattt tagtcatatc agataagcat tgattaatat
     4681 cattattgct tctacaagct ttaattttat taattattct gtatgtgtcg tcggcattta
     4741 tgtttttcat attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     4801 gaaaggtttt gcgccattcg atggcgcgcc gcttcgtcag gccacatagc tttcttgttc
     4861 tgatcggaac gatcgttggc tgacctgtag gatcgtacag gtttacgcaa gaaaatggtt
     4921 tgttatagtc gaataaatac tagagaaaga ggagaaatac cat
//
