LOCUS       sequence-74541-e        5542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5542)
  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 5542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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: pVRc14_1324
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1027..1044)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1197..1283
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1375..1402
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1421..1512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1682..2341
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1748..1767)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2483..2938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2570..2589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3624..3641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3758..4303)
                     /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(3838..3857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4375..4394
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5525..5536
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgagcggta gccgtccgga accggaaggt catctggcag aacagcatct gggtgatcgt
       61 ctgagcgcac tggttgatgg tgaactgggt catgatgcac gtgaacgtgt tctggcacat
      121 gttgcaacct gtccgaaatg taaagccgaa gttgatgcac agcgtcgtct gaaaaatgtt
      181 tttgcagaag cagcaccgcc tgcaccgagc gaaagctttc tggctcgtct gcagggtctg
      241 cctggtggtg gtgatagtga tggcggtggt agcccgttta gcggtctgcc aggcggtttt
      301 ggtgcaagcg cagcaagcgg tgtttttggt ccgcgtcgtg atgaacgttt tgagtttgat
      361 tatgttccgg caggtagcca tacaccggtt ctgccgagcg caaccagcgg tcgtggtttt
      421 cgtattcatg aagttggtcg tcatgaaagc gatcgtagcg caagccgtgg tctgcgtttt
      481 gcatttgcag cagccggtgc agttagcctg gcagcaattg cactgggtgg tgttaccctg
      541 ggtacaccgg ataccaccac cgaagcacgt ggtagcggta gtggtagcaa tgttacaccg
      601 ctgcgcacac cgggttcagc agcagcaacc ggtagcgaaa gtcagcgtcg ccgtaccgca
      661 ggtccgctgc tgggtcaggg tcaacgtgcc ctgggcgatc tgccggttgc aagcaccacc
      721 gcaagtgcac ctctgctgcc aggtatgcca gcaccggcag gcggtgatgc ccgtcagcag
      781 gcagttcgtg cactgaccac accggtgacc gcaggcgcag ccgcaatgag tccgctgatt
      841 cgtccgctgg aagcagttcc gcctctgagc ctgagcagct ggtcagccgc accggaagtt
      901 cgtcctccgg gtctgctggc agcccctgat ccggcaccga gtccgtatcc ggcagccagc
      961 cctgccgcaa gcagctctcc gctgcgttaa ggatccctgt tttggcggat gagagaagat
     1021 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     1081 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     1141 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     1201 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1261 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     1321 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     1381 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
     1441 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1501 aaaaggaaga ctatgagtat tcaacatttc cgtgtctgct tacataaaca gtaatacaag
     1561 gggtgtttac tagaggttga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     1621 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1681 aatggagaaa aaaatcacgg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1741 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1801 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1861 tcacattctt gcccgcctga tgaacgctca cccggagttt cgtatggcca tgaaagacgg
     1921 tgagctggtg atctgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     1981 aacgttttcg tccctctgga gtgaatacca cgacgatttc cggcagtttc tccacatata
     2041 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2101 gaatatgttt tttgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2161 ggccaatatg gacaacttct tcgcccccgt tttcacgatg ggcaaatatt atacgcaagg
     2221 cgacaaggtg ctgatgccgc tggcgatcca ggttcatcat gccgtttgtg atggcttcca
     2281 tgtcggccgc atgcttaatg aattacaaca gtactgtgat gagtggcagg gcggggcgta
     2341 ataatactag ctccggcaaa aaaacgggca aggtgtcacc accctgccct ttttctttaa
     2401 aaccgaaaag attacttcgc gtttgccacc tgacgtctaa gctgtcagac caagtttact
     2461 catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2761 gagtccacgt tctttaatag tggactcttg ttccaaactt gaacaacact caaccctatc
     2821 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2881 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttaa
     2941 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3001 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3181 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtcaggcatt tgagaagcac
     3301 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     3361 atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat
     3421 tcatccgctt attatcactt attcaggcgt agcaccaggc gtttaagggc accaataact
     3481 gccttaaaaa aagctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     3541 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     3601 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     3661 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     3721 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     3781 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     3841 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     3901 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     3961 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     4021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     4081 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     4141 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     4201 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     4261 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     4321 tcagataaaa tatttgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     4381 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     4441 tccggcgtag aggatctgct catgtttgac agcttatcat cgatgcataa tgtgcctgtc
     4501 aaatggacga agcagggatt ctgcaaaccc tatgctactc cctcgagccg tcaattgtct
     4561 gattcgttac caattattaa tttttaaagt atgggcaatc aattgctcct gttaaaattg
     4621 ctttagaaat actttggcag cggtttgttg tattgagttt catttgcgca ttggttaaat
     4681 ggaaagtgac agtacgctca ctgcaaccta atatttttga aatatcccaa gagctttttc
     4741 cttcgcatgc ccacgctaaa cattcttttt ctcttttggt taaatcgttg tttgatttat
     4801 tatttgctat atttattttt cgataattat caactagaga aggaacaatt aatggtatgt
     4861 tcatacacgc atgtaaaaat aaactatcta tatagttgtc tttttctgaa tgtgcaaaac
     4921 taagcattcc gaagccattg ttagccgtat gaatagggaa actaaaccca gtgataagac
     4981 ctgatgtttt cgcttcttta attacatttg gagatttttt atttacagca ttgttttcaa
     5041 atatattcca attaattggt gaatgattgg agttagaata atctactata ggatcatatt
     5101 ttattaaatt agcgtcatca taatattgcc tccatttttt agggtaatta tctaggattg
     5161 aaatatcaga tttaaccata gaatgaggat aaatgatcgc gagtaaataa tattcacaat
     5221 gtaccatttt agtcatatca gataagcatt gattaatatc attattgctt ctacaagctt
     5281 taattttatt aattattctg tatgtgtcgt cggcatttat gtttttcata ttcaccaccc
     5341 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     5401 tggcgcgccg cttcgtcagg ccacatagct ttcttgttct gatcggaacg atcgttggct
     5461 gacctgtagg atcgtacagg tttacgcaag aaaatggttt gttatagtcg aataaatact
     5521 agagaaagag gagaaatacc at
//
