LOCUS       sequence-74546-e        4984 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 4984)
  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 4984)
  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: pVRc20_992
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(469..486)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      639..725
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      817..844
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        863..954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1124..1783
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1190..1209)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1925..2380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2012..2031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3066..3083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3200..3745)
                     /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(3280..3299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3817..3836
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4967..4978
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgacaccgg aacgttttgt tcatctggca gatgcctatg gtgccgatct gcagcgttgg
       61 ccgagcgcag aacgtgcagc agcacaggca ctgctggatt gtggtgatgc acaggcagtt
      121 gcagcactgc gtcaggcaca ttggctggat agccagctgg atcgttatca ggttccggca
      181 ccgagtccgg cactggcaca gcgtattatt gcagcagccc agcagcctgg tgcaccgttt
      241 tggagccgtt atgcaggttg gctggcaagc ctgggttggg ttggtgttgg tctgaccggt
      301 gttgcagcag gtatgctggc agttgcactg agcctgccgc tgagcaccag cgcagaagca
      361 ctgccgagcg tttttgatca gagtgatgca gaatttgtgc tgagcattaa tgcagaagaa
      421 gcagaacagt aaggatccct gttttggcgg atgagagaag attttcagcc tgatacagat
      481 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
      541 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
      601 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      661 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      721 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
      781 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
      841 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
      901 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gactatgagt
      961 attcaacatt tccgtgtctg cttacataaa cagtaataca aggggtgttt actagaggtt
     1021 gatcgggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     1081 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     1141 gggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     1201 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     1261 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     1321 gatgaacgct cacccggagt ttcgtatggc catgaaagac ggtgagctgg tgatctggga
     1381 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt cgtccctctg
     1441 gagtgaatac cacgacgatt tccggcagtt tctccacata tattcgcaag atgtggcgtg
     1501 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt tttttgtctc
     1561 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     1621 cttcgccccc gttttcacga tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     1681 gctggcgatc caggttcatc atgccgtttg tgatggcttc catgtcggcc gcatgcttaa
     1741 tgaattacaa cagtactgtg atgagtggca gggcggggcg taataatact agctccggca
     1801 aaaaaacggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc
     1861 gcgtttgcca cctgacgtct aagctgtcag accaagttta ctcatatata ctttagattg
     1921 atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1981 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2041 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2101 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2161 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2221 agtggactct tgttccaaac ttgaacaaca ctcaacccta tctcgggcta ttcttttgat
     2281 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2341 tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat
     2401 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2461 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2521 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2581 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2641 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2701 cgctctgcta atcctgttac cagtcaggca tttgagaagc acacggtcac actgcttccg
     2761 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     2821 gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
     2881 ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaagctagc
     2941 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     3001 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     3061 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     3121 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     3181 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     3241 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3301 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     3361 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     3421 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     3481 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     3541 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     3601 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     3661 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     3721 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttgct
     3781 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     3841 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     3901 ctcatgtttg acagcttatc atcgatgcat aatgtgcctg tcaaatggac gaagcaggga
     3961 ttctgcaaac cctatgctac tccctcgagc cgtcaattgt ctgattcgtt accaattatt
     4021 aatttttaaa gtatgggcaa tcaattgctc ctgttaaaat tgctttagaa atactttggc
     4081 agcggtttgt tgtattgagt ttcatttgcg cattggttaa atggaaagtg acagtacgct
     4141 cactgcaacc taatattttt gaaatatccc aagagctttt tccttcgcat gcccacgcta
     4201 aacattcttt ttctcttttg gttaaatcgt tgtttgattt attatttgct atatttattt
     4261 ttcgataatt atcaactaga gaaggaacaa ttaatggtat gttcatacac gcatgtaaaa
     4321 ataaactatc tatatagttg tctttttctg aatgtgcaaa actaagcatt ccgaagccat
     4381 tgttagccgt atgaataggg aaactaaacc cagtgataag acctgatgtt ttcgcttctt
     4441 taattacatt tggagatttt ttatttacag cattgttttc aaatatattc caattaattg
     4501 gtgaatgatt ggagttagaa taatctacta taggatcata ttttattaaa ttagcgtcat
     4561 cataatattg cctccatttt ttagggtaat tatctaggat tgaaatatca gatttaacca
     4621 tagaatgagg ataaatgatc gcgagtaaat aatattcaca atgtaccatt ttagtcatat
     4681 cagataagca ttgattaata tcattattgc ttctacaagc tttaatttta ttaattattc
     4741 tgtatgtgtc gtcggcattt atgtttttca tattcaccac cctgaattga ctctcttccg
     4801 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggcgcgc cgcttcgtca
     4861 ggccacatag ctttcttgtt ctgatcggaa cgatcgttgg ctgacctgta ggatcgtaca
     4921 ggtttacgca agaaaatggt ttgttatagt cgaataaata ctagagaaag aggagaaata
     4981 ccat
//
