LOCUS       sequence-74556-e        5386 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 5386)
  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 5386)
  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: pVRc33_423
FEATURES             Location/Qualifiers
     source          1..5386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(871..888)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1041..1127
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1219..1246
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1265..1356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1526..2185
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1592..1611)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2327..2782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2414..2433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3468..3485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3602..4147)
                     /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(3682..3701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4219..4238
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5369..5380
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgatggcaa tgagcaatac catgcctgat ccgcgtgaac cgggtgatgt tgaagcactg
       61 ctgccgtggt atgcagcagg caccctgaat gcacgtgatg cacgtcgtgt tgcagatgca
      121 ctggatcgtg atccggcact ggcacgtcag tatgcagtta ttctggaaga atatgcaagc
      181 accattgaac tgaatgaaag cctgggtgca ccgagcagcc gtgcaatgca gaaactgttt
      241 gcagcaattg atgcagaacc ggcacgtgca ccgggtgcag gtcagggtct gggtgcacgt
      301 tttgcaggtt tttttgcagg tctgagcccg aaagcactgg catggtcagc aagcgttgca
      361 ggtctggcac tgctgctgca ggcaggtctg attggtgcac tgctgacctg gcctcatgca
      421 gcaccggttc agaccgcagc atatcagccg cagcgtgaag ttgcccgtgc accggcaagt
      481 agtccggcac cggcaaccgt tagccctccg gcagcagcaa tggcagatcg tgcagcagat
      541 gcaggtaaaa gcacccctcc gatggttatg gcagaacgta gcggtggtcc ggttgttcgt
      601 agcctggcac cgcagagcgg tccgcgtgtt ctggttaaat ttgcaccgga agcacgtgca
      661 agcgaaattg cagcactgct ggatcagtat aatgcagttg ttgttgatag cagccgtggt
      721 ggtctgtttc gtctgcagtt tggcacccag agcctgagca aacaggatca agaaaccctg
      781 attggtcgtc tgcagaaaga accggttgtt agcgttgttc tgagcgcacc gtaaggatcc
      841 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
      901 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
      961 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     1021 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1081 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     1141 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     1201 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     1261 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1321 tgataaatgc ttcaataata ttgaaaaagg aagactatga gtattcaaca tttccgtgtc
     1381 tgcttacata aacagtaata caaggggtgt ttactagagg ttgatcgggc acgtaagagg
     1441 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     1501 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc acgggatata ccaccgttga
     1561 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1621 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1681 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaacg ctcacccgga
     1741 gtttcgtatg gccatgaaag acggtgagct ggtgatctgg gatagtgttc acccttgtta
     1801 caccgttttc catgagcaaa ctgaaacgtt ttcgtccctc tggagtgaat accacgacga
     1861 tttccggcag tttctccaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1921 ctatttccct aaagggttta ttgagaatat gttttttgtc tcagccaatc cctgggtgag
     1981 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     2041 gatgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga tccaggttca
     2101 tcatgccgtt tgtgatggct tccatgtcgg ccgcatgctt aatgaattac aacagtactg
     2161 tgatgagtgg cagggcgggg cgtaataata ctagctccgg caaaaaaacg ggcaaggtgt
     2221 caccaccctg ccctttttct ttaaaaccga aaagattact tcgcgtttgc cacctgacgt
     2281 ctaagctgtc agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc
     2341 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2401 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2461 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2521 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2581 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2641 acttgaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     2701 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2761 aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2821 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2881 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2941 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3001 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3061 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3121 accagtcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     3181 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     3241 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
     3301 aggcgtttaa gggcaccaat aactgcctta aaaaaagcta gcggagtgta tactggctta
     3361 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     3421 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     3481 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     3541 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     3601 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     3661 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     3721 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     3781 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     3841 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3901 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     3961 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     4021 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     4081 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     4141 atctcaagaa gatcatctta ttaatcagat aaaatatttg ctcatgagcc cgaagtggcg
     4201 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     4261 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     4321 tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
     4381 actccctcga gccgtcaatt gtctgattcg ttaccaatta ttaattttta aagtatgggc
     4441 aatcaattgc tcctgttaaa attgctttag aaatactttg gcagcggttt gttgtattga
     4501 gtttcatttg cgcattggtt aaatggaaag tgacagtacg ctcactgcaa cctaatattt
     4561 ttgaaatatc ccaagagctt tttccttcgc atgcccacgc taaacattct ttttctcttt
     4621 tggttaaatc gttgtttgat ttattatttg ctatatttat ttttcgataa ttatcaacta
     4681 gagaaggaac aattaatggt atgttcatac acgcatgtaa aaataaacta tctatatagt
     4741 tgtctttttc tgaatgtgca aaactaagca ttccgaagcc attgttagcc gtatgaatag
     4801 ggaaactaaa cccagtgata agacctgatg ttttcgcttc tttaattaca tttggagatt
     4861 ttttatttac agcattgttt tcaaatatat tccaattaat tggtgaatga ttggagttag
     4921 aataatctac tataggatca tattttatta aattagcgtc atcataatat tgcctccatt
     4981 ttttagggta attatctagg attgaaatat cagatttaac catagaatga ggataaatga
     5041 tcgcgagtaa ataatattca caatgtacca ttttagtcat atcagataag cattgattaa
     5101 tatcattatt gcttctacaa gctttaattt tattaattat tctgtatgtg tcgtcggcat
     5161 ttatgttttt catattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5221 cgcgaaaggt tttgcgccat tcgatggcgc gccgcttcgt caggccacat agctttcttg
     5281 ttctgatcgg aacgatcgtt ggctgacctg taggatcgta caggtttacg caagaaaatg
     5341 gtttgttata gtcgaataaa tactagagaa agaggagaaa taccat
//
