LOCUS       sequence-74544-e        5194 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 5194)
  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 5194)
  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: pVRc16_973
FEATURES             Location/Qualifiers
     source          1..5194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(679..696)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      849..935
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1027..1054
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1073..1164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1334..1993
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1400..1419)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2135..2590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2222..2241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3276..3293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3410..3955)
                     /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(3490..3509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4027..4046
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5177..5188
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaaaaccg atgatctgat tgcactgctg gcagccggtg aaggtccggt tccgcgtcat
       61 gcagttggtc gtcgtatggc agttgcagca ctgggtggtc tgaccgcagc actgctgctg
      121 accattaccc tgtatggtgt tcgtagcgat attaccgaag ttgcacagac accgctgttt
      181 tggggtaaag ttgcatttcc gaccagcctg gcactgattg gtctgtggct gaccagccgt
      241 ctggcacgtc cgggtggtaa aggtgcagca ggttggaaaa tgctgggtct gccgctgctg
      301 ctggtttggt gtggtgcagc agttagcatt gcgggtgcac cggttgatgc acgtgcagat
      361 ctgctgtttg gtcgtacctg gcgtacctgt gcactgaata ttgccctgct gagcgttccg
      421 gcatttgtta ccgttttttg ggcactgaaa ggtctggcac cgacccgtct gcgtctggct
      481 ggcgcagccg gtggtctgct ggcaggtagc agcgcaaccg ttgcatattg tctgcattgt
      541 ccggaaatgg gtgttccgtt ttggggtgtt tggtatgttc tgggtatgct ggttccgacc
      601 gttctgggtg catggtgggg tccgcgtatg ctgcgttggt aaggatccct gttttggcgg
      661 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
      721 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
      781 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
      841 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      901 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
      961 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     1021 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta
     1081 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1141 caataatatt gaaaaaggaa gactatgagt attcaacatt tccgtgtctg cttacataaa
     1201 cagtaataca aggggtgttt actagaggtt gatcgggcac gtaagaggtt ccaactttca
     1261 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
     1321 taaggaagct aaaatggaga aaaaaatcac gggatatacc accgttgata tatcccaatg
     1381 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     1441 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     1501 tccggccttt attcacattc ttgcccgcct gatgaacgct cacccggagt ttcgtatggc
     1561 catgaaagac ggtgagctgg tgatctggga tagtgttcac ccttgttaca ccgttttcca
     1621 tgagcaaact gaaacgtttt cgtccctctg gagtgaatac cacgacgatt tccggcagtt
     1681 tctccacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     1741 agggtttatt gagaatatgt tttttgtctc agccaatccc tgggtgagtt tcaccagttt
     1801 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacga tgggcaaata
     1861 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatc caggttcatc atgccgtttg
     1921 tgatggcttc catgtcggcc gcatgcttaa tgaattacaa cagtactgtg atgagtggca
     1981 gggcggggcg taataatact agctccggca aaaaaacggg caaggtgtca ccaccctgcc
     2041 ctttttcttt aaaaccgaaa agattacttc gcgtttgcca cctgacgtct aagctgtcag
     2101 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     2161 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2221 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2281 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2341 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2401 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac ttgaacaaca
     2461 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2521 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2581 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2821 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtcaggca
     2941 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     3001 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     3061 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg
     3121 gcaccaataa ctgccttaaa aaaagctagc ggagtgtata ctggcttact atgttggcac
     3181 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     3241 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     3301 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     3361 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     3421 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     3481 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     3541 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     3601 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     3661 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     3721 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     3781 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     3841 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     3901 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     3961 tcatcttatt aatcagataa aatatttgct catgagcccg aagtggcgag cccgatcttc
     4021 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     4081 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc atcgatgcat
     4141 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccctcgagc
     4201 cgtcaattgt ctgattcgtt accaattatt aatttttaaa gtatgggcaa tcaattgctc
     4261 ctgttaaaat tgctttagaa atactttggc agcggtttgt tgtattgagt ttcatttgcg
     4321 cattggttaa atggaaagtg acagtacgct cactgcaacc taatattttt gaaatatccc
     4381 aagagctttt tccttcgcat gcccacgcta aacattcttt ttctcttttg gttaaatcgt
     4441 tgtttgattt attatttgct atatttattt ttcgataatt atcaactaga gaaggaacaa
     4501 ttaatggtat gttcatacac gcatgtaaaa ataaactatc tatatagttg tctttttctg
     4561 aatgtgcaaa actaagcatt ccgaagccat tgttagccgt atgaataggg aaactaaacc
     4621 cagtgataag acctgatgtt ttcgcttctt taattacatt tggagatttt ttatttacag
     4681 cattgttttc aaatatattc caattaattg gtgaatgatt ggagttagaa taatctacta
     4741 taggatcata ttttattaaa ttagcgtcat cataatattg cctccatttt ttagggtaat
     4801 tatctaggat tgaaatatca gatttaacca tagaatgagg ataaatgatc gcgagtaaat
     4861 aatattcaca atgtaccatt ttagtcatat cagataagca ttgattaata tcattattgc
     4921 ttctacaagc tttaatttta ttaattattc tgtatgtgtc gtcggcattt atgtttttca
     4981 tattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5041 tgcgccattc gatggcgcgc cgcttcgtca ggccacatag ctttcttgtt ctgatcggaa
     5101 cgatcgttgg ctgacctgta ggatcgtaca ggtttacgca agaaaatggt ttgttatagt
     5161 cgaataaata ctagagaaag aggagaaata ccat
//
