LOCUS       sequence-97574-e        8022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses T7 RNAP from the arabinose inducible promoter PBAD with a 
            RBS strength of 500 from the Salis RBS calculator. Also includes a 
            constitutive araC ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8022)
  AUTHORS   Shis DL, Bennett MR
  TITLE     Library of synthetic transcriptional AND gates built with split T7 
            RNA polymerase mutants.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5028-33. doi: 
            10.1073/pnas.1220157110. Epub 2013 Mar 11.
  PUBMED    23479654
REFERENCE   2  (bases 1 to 8022)
  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"; doi: 
            "10.1073/pnas.1220157110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-03-26- 26"; volume: "110"; issue: "13"; pages: "5028-33"
COMMENT     Sequence Label: pTara:500
FEATURES             Location/Qualifiers
     source          1..8022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1289
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1212..1231
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(4065..4082)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4235..4321
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4413..4440
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4459..4550
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5004..5459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5091..5110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(6014..6673)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6588..6607
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6674..6776)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7168..7185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7302..7847)
                     /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(7382..7401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7919..7938
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtcctaggac attgattatt tgcacggcgt
     1201 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     1261 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagcg agctctagaa
     1321 agacttactt aataacattc tttttctaat atgaacacga ttaacatcgc taagaacgac
     1381 ttctctgaca tcgaactggc tgctatcccg ttcaacactc tggctgacca ttacggtgag
     1441 cgtttagctc gcgaacagtt ggcccttgag catgagtctt acgagatggg tgaagcacgc
     1501 ttccgcaaga tgtttgagcg tcaacttaaa gctggtgagg ttgcggataa cgctgccgcc
     1561 aagcctctca tcactaccct actccctaag atgattgcac gcatcaacga ctggtttgag
     1621 gaagtgaaag ctaagcgcgg caagcgcccg acagccttcc agttcctgca agaaatcaag
     1681 ccggaagccg tagcgtacat caccattaag accactctgg cttgcctaac cagtgctgac
     1741 aatacaaccg ttcaggctgt agcaagcgca atcggtcggg ccattgagga cgaggctcgc
     1801 ttcggtcgta tccgtgacct tgaagctaag cacttcaaga aaaacgttga ggaacaactc
     1861 aacaagcgcg tagggcacgt ctacaagaaa gcatttatgc aagttgtcga ggctgacatg
     1921 ctctctaagg gtctactcgg tggcgaggcg tggtcttcgt ggcataagga agactctatt
     1981 catgtaggag tacgctgcat cgagatgctc attgagtcaa ccggaatggt tagcttacac
     2041 cgccaaaatg ctggcgtagt aggtcaagac tctgagacta tcgaactcgc acctgaatac
     2101 gctgaggcta tcgcaacccg tgcaggtgcg ctggctggca tctctccgat gttccaacct
     2161 tgcgtagttc ctcctaagcc gtggactggc attactggtg gtggctattg ggctaacggt
     2221 cgtcgtcctc tggcgctggt gcgtactcac agtaagaaag cactgatgcg ctacgaagac
     2281 gtttacatgc ctgaggtgta caaagcgatt aacattgcgc aaaacaccgc atggaaaatc
     2341 aacaagaaag tcctagcggt cgccaacgta atcaccaagt ggaagcattg tccggtcgag
     2401 gacatccctg cgattgagcg tgaagaactc ccgatgaaac cggaagacat cgacatgaat
     2461 cctgaggctc tcaccgcgtg gaaacgtgct gccgctgctg tgtaccgcaa ggacaaggct
     2521 cgcaagtctc gccgtatcag ccttgagttc atgcttgagc aagccaataa gtttgctaac
     2581 cataaggcca tctggttccc ttacaacatg gactggcgcg gtcgtgttta cgctgtgtca
     2641 atgttcaacc cgcaaggtaa cgatatgacc aaaggactgc ttacgctggc gaaaggtaaa
     2701 ccaatcggta aggaaggtta ctactggctg aaaatccacg gtgcaaactg tgcgggtgtc
     2761 gataaggttc cgttccctga gcgcatcaag ttcattgagg aaaaccacga gaacatcatg
     2821 gcttgcgcta agtctccact ggagaacact tggtgggctg agcaagattc tccgttctgc
     2881 ttccttgcgt tctgctttga gtacgctggg gtacagcacc acggcctgag ctataactgc
     2941 tcccttccgc tggcgtttga cgggtcttgc tctggcatcc agcacttctc cgcgatgctc
     3001 cgagatgagg taggtggtcg cgcggttaac ttgcttccta gtgaaaccgt tcaggacatc
     3061 tacgggattg ttgctaagaa agtcaacgag attctacaag cagacgcaat caatgggacc
     3121 gataacgaag tagttaccgt gaccgatgag aacactggtg aaatctctga gaaagtcaag
     3181 ctgggcacta aggcactggc tggtcaatgg ctggcttacg gtgttactcg cagtgtgact
     3241 aagcgttcag tcatgacgct ggcttacggg tccaaagagt tcggcttccg tcaacaagtg
     3301 ctggaagata ccattcagcc agctattgat tccggcaagg gtctgatgtt cactcagccg
     3361 aatcaggctg ctggatacat ggctaagctg atttgggaat ctgtgagcgt gacggtggta
     3421 gctgcggttg aagcaatgaa ctggcttaag tctgctgcta agctgctggc tgctgaggtc
     3481 aaagataaga agactggaga gattcttcgc aagcgttgcg ctgtgcattg ggtaactcct
     3541 gatggtttcc ctgtgtggca ggaatacaag aagcctattc agacgcgctt gaacctgatg
     3601 ttcctcggtc agttccgctt acagcctacc attaacacca acaaagatag cgagattgat
     3661 gcacacaaac aggagtctgg tatcgctcct aactttgtac acagccaaga cggtagccac
     3721 cttcgtaaga ctgtagtgtg ggcacacgag aagtacggaa tcgaatcttt tgcactgatt
     3781 cacgactcct tcggtaccat tccggctgac gctgcgaacc tgttcaaagc agtgcgcgaa
     3841 actatggttg acacatatga gtcttgtgat gtactggctg atttctacga ccagttcgct
     3901 gaccagttgc acgagtctca attggacaaa atgccagcac ttccggctaa aggtaacttg
     3961 aacctccgtg acatcttaga gtcggacttc gcgttcgcgt aatctagagt cgacctgcag
     4021 gcatgcaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     4081 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     4141 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     4201 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     4261 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     4321 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     4381 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     4441 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttggggcaa actattaact
     4681 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4741 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4801 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4861 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4921 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4981 tcatatatac tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5041 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5101 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5161 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5221 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5281 ggagtccacg ttctttaata gtggactctt gttccaaact tgaacaacac tcaaccctat
     5341 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5401 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta
     5461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca
     5821 cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc
     5881 tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca
     5941 ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg caccaataac
     6001 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     6061 gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca
     6121 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     6181 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     6241 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     6301 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     6361 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     6421 atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc
     6481 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     6541 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     6601 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     6661 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     6721 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     6781 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     6841 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     6901 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     6961 gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
     7021 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
     7081 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     7141 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     7201 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     7261 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     7321 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     7381 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     7441 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     7501 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     7561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     7621 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     7681 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     7741 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     7801 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     7861 ttaatcagat aaaatatttg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     7921 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     7981 tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tc
//
