LOCUS       62056_1540        5228 bp DNA     circular SYN 03-DEC-2019
DEFINITION  Cloning vector Master_Architecture_RbsR_HQN, complete sequence.
ACCESSION   MN207947
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5228)
  AUTHORS   Rondon RE, Groseclose TM, Short AE, Wilson CJ.
  TITLE     Transcriptional programming using engineered systems of 
            transcription factors and genetic architectures
  JOURNAL   Nat Commun 10 (1), 4784 (2019)
  PUBMED    31636266
REFERENCE   2  (bases 1 to 5228)
  AUTHORS   Rondon RE, Groseclose T, Short A, Wilson CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUL-2019) Chemical and Biomolecular Engineering, 
            Georgia Institute of Technology, 950 Atlantic dr. NW, 5110E, 
            Atlanta, GA 30332, United States
REFERENCE   3  (bases 1 to 5228)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2019"; volume: "10"; issue: "1"; pages: "4784"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-JUL-2019) Chemical and Biomolecular Engineering, Georgia 
            Institute of Technology, 950 Atlantic dr. NW, 5110E, Atlanta, GA 
            30332, United States"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      51..56
                     /label=trc promoter -35 hexamer TTGACA
                     /note="trc promoter -35 hexamer TTGACA"
                     /regulatory_class="promoter"
     regulatory      57..73
                     /label=operator Ottg
                     /note="operator Ottg"
                     /regulatory_class="other"
     regulatory      74..79
                     /label=trc promoter -10 hexamer TATAAT
                     /note="trc promoter -10 hexamer TATAAT"
                     /regulatory_class="promoter"
     regulatory      95..114
                     /label=operator Oagg
                     /note="operator Oagg"
                     /regulatory_class="other"
     regulatory      133..182
                     /label=RiboJ10
                     /note="RiboJ10"
                     /regulatory_class="insulator"
     regulatory      224..235
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             242..955
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     terminator      970..1056
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(1261..2208)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(2256..2478)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     regulatory      2565..2570
                     /label=pLL promoter -35 hexamer TTGACA
                     /note="pLL promoter -35 hexamer TTGACA"
                     /regulatory_class="promoter"
     regulatory      2571..2587
                     /label=operator Osym
                     /note="operator Osym"
                     /regulatory_class="other"
     regulatory      2588..2593
                     /label=pLL promoter -10 hexamer GATACT
                     /note="pLL promoter -10 hexamer GATACT"
                     /regulatory_class="promoter"
     regulatory      2609..2628
                     /label=operator Otta
                     /note="operator Otta"
                     /regulatory_class="other"
     misc_RNA        2646..2696
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      2698..2719
                     /label=RiboJ
                     /note="RiboJ"
                     /regulatory_class="insulator"
     regulatory      2724..2735
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             2742..3740
                     /codon_start=1
                     /transl_table=11
                     /product="RbsR HQN"
                     /label=RbsR HQN
                     /protein_id="QFU95565.1"
                     /translation="MKPVTLYDVAEYAGVSHQTVSNVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKASHTIGMLITASTNPFYSELVRGVERSCFERGYSLVLCNTEGDEQ
                     RMNRNLETLMQKRVDGLLLLCTETHQPSREIMQRYPTVPTVMMDWAPFDGDSDLIQDNS
                     LLGGDLATQYLIDKGHTRIACITGPLDKTPARLRLEGYRAAMKRAGLNIPDGYEVTGDF
                     EFNGGFDAMRQLLSHPLRPQAVFTGNDAMAVGVYQALYQAELQVPQDIAVIGYDDIELA
                     SFMTPPLTTIHQPKDELGELAIDVLIHRITQPTLQQQRLQLTPILMERGSA"
     terminator      3741..3772
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     terminator      complement(4173..4267)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4301..5092)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gacgtccaac atatggcacc ggaggctttc gtcttcacct cgagaattgt ttgacatttt
       61 gagcgctcaa aattataatt cgtgcacttc gtgcaattag gagcgctcct aattggactg
      121 cttgcacctg agcgctcaac gggtgtgctt cccgttctga tgagtccgtg aggacgaaag
      181 cgcctctaca aataattttg tttaactgag cacagaattc attaaagagg agaaaggtac
      241 catgagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
      301 tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg ctacaaacgg
      361 aaagcttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
      421 tgtcactact ctcacttatg gtgttcaatg cttttcccgt tatccggatc atatgaaacg
      481 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
      541 caaagatgac gggacttaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
      601 taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa
      661 acttgagtac aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
      721 aatcaaagct aacttcaaaa ttcgccacaa cgttgaagat ggatccgttc aactagcaga
      781 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
      841 cctgtcgaca caatctgtcc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct
      901 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct acaaataatc
      961 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1021 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggtacg
     1081 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     1141 tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt
     1201 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagctca
     1261 gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca
     1321 tgagaacgaa ccattgagat catgcttact ttgcatgtca ctcaaaaatt ttgcctcaaa
     1381 actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttacg
     1441 taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt
     1501 gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt
     1561 atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc
     1621 tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc
     1681 aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt
     1741 cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa
     1801 agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa
     1861 tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca
     1921 ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag
     1981 ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg
     2041 agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt
     2101 ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata
     2161 gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg
     2221 gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt
     2281 ccttttcccg ggagatctgg gtatctgtaa attctgctag acctttgctg gaaaacttgt
     2341 aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat
     2401 tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc
     2461 cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca
     2521 aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaagttgaca ttgtgagcgc
     2581 tcacaatgat actatatagt gaacaaggaa ttttaagcgc ttaaaattga ttcctgttag
     2641 gatcagctgt caccggatgt gctttccggt ctgatgagtc cgtgaggacg aaacagcctc
     2701 tacaaataat tttgtttaac ctcaaagagg agaaactcga cgtgaaacca gtaacgttat
     2761 acgatgtcgc agagtatgcc ggtgtctctc atcagaccgt ttccaatgtg gtgaaccagg
     2821 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctcaatt
     2881 acattcccaa ccgcgtggca caacaactgg cgggcaaagc gagccatacc attggcatgt
     2941 tgatcactgc cagtaccaat cctttctatt cagaactggt gcgtggcgtt gaacgcagct
     3001 gcttcgaacg cggttatagt ctcgtccttt gcaataccga aggcgatgaa cagcggatga
     3061 atcgcaatct ggaaacgctg atgcaaaaac gcgttgatgg cttgctgtta ctgtgcaccg
     3121 aaacgcatca accttcgcgt gaaatcatgc aacgttatcc gacagtgcct actgtgatga
     3181 tggactgggc tccgttcgat ggcgacagcg atcttattca ggataactcg ttgctgggcg
     3241 gagacttagc aacgcaatat ctgatcgata aaggtcatac ccgtatcgcc tgtattaccg
     3301 gcccgctgga taaaactccg gcgcgcctgc ggttggaagg ttatcgggcg gcgatgaaac
     3361 gtgcgggtct caacattcct gatggctatg aagtcactgg tgattttgaa tttaacggcg
     3421 ggtttgacgc tatgcgccaa ctgctatcac atccgctgcg tcctcaggcc gtctttaccg
     3481 gaaatgacgc tatggctgtt ggcgtttacc aggcgttata tcaggcagag ttacaggttc
     3541 cgcaggatat cgcggtgatt ggctatgacg atatcgaact ggcaagcttt atgacgccac
     3601 cattaaccac tatccaccaa ccgaaagatg aactggggga gctggcgatt gatgtactca
     3661 tccatcggat aacccagccg acccttcagc aacaacgatt acaacttact ccgattctga
     3721 tggaacgcgg ttcggcttag agtcaaaagc ctccgaccgg aggcttttga ctagtagcac
     3781 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     3841 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     3901 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     3961 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     4021 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     4081 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     4141 ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     4201 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     4261 aggagtccaa gcgagctctc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     4321 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     4381 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     4441 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     4501 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     4561 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     4621 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     4681 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     4741 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     4801 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     4861 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     4921 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     4981 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     5041 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     5101 atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca
     5161 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     5221 gcaattcc
//
