LOCUS       40924_4160        5584 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAH12, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  AUTHORS   Kelly CL, Harris AW, Steel H, Hancock EJ, Heap JT, Papachristodoulou
            A.
  TITLE     Synthetic negative feedback circuits using engineered small RNAs
  JOURNAL   Nucleic Acids Res. (2018) In press
  PUBMED    30212900
REFERENCE   2  (bases 1 to 5584)
  AUTHORS   Kelly CL, Harris AWK., Steel H, Hancock EJ, Heap JT, 
            Papachristodoulou A.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-SEP-2018) School of Natural and Environmental 
            Sciences, Newcastle University, Agriculture Building, King's Road, 
            Newcastle NE1 7RU, United Kingdom
REFERENCE   3  (bases 1 to 5584)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5584)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-SEP-2018) School of Natural and Environmental Sciences, 
            Newcastle University, Agriculture Building, King's Road, Newcastle 
            NE1 7RU, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     875..891
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(908..1021)
                     /label=rpn txn term terminator
                     /note="rpn txn term terminator"
                     /regulatory_class="terminator"
     regulatory      complement(1028..1328)
                     /label=bla txn term terminator
                     /note="bla txn term terminator"
                     /regulatory_class="terminator"
     regulatory      1346..1489
                     /label=proD promoter
                     /note="proD promoter"
                     /regulatory_class="promoter"
     regulatory      1490..1523
                     /label=Ecoli rhaS RBS
                     /note="Ecoli rhaS RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1524..2357
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     CDS             2358..3068
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     CDS             3087..3104
                     /label=6xHis
                     /note="6xHis affinity tag"
     regulatory      3137..3311
                     /label=rrnB1 B2 T1 term terminator
                     /note="rrnB1 B2 T1 term terminator"
                     /regulatory_class="terminator"
     promoter        3398..3489
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3420..3448
                     /label=ampR promoter
                     /note="ampR promoter"
                     /regulatory_class="promoter"
     CDS             3490..4347
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             4433..5512
                     /label=lacI
                     /note="lac repressor"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttagccca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaaggcga gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg
      841 gcctttttgc gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc
      901 gggccccctg ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc
      961 gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa
     1021 gggcgcctgt cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag
     1081 caacgcactt taaataagat acgttgcttt ttcgattgat gaacacctat aattaaacta
     1141 ttcatctatt atttatgatt ttttgtatat acaatatttc tagtttgtta aagagaatta
     1201 agaaaataaa tctcgaaaat aataaaggga aaatcagttt ttgatatcaa aattatacat
     1261 gtcaacgata atacaaaata taatacaaac tataagatgt tatcagtatt tattatgcat
     1321 ttagaataaa ttttgtgtcg cccttcacag ctaacaccac gtcgtcccta tctgctgccc
     1381 taggtctatg agtggttgct ggataacttt acgggcatgc ataaggctcg tataatatat
     1441 tcagggagac cacaacggtt tccctctaca aataattttg tttaactttt atttcgccgt
     1501 gttgacgaca tcaggaggcc agtatgaccg tattacatag tgtggatttt tttccgtctg
     1561 gtaacgcgtc cgtggcgata gaaccccggc tcccgcaggc ggattttcct gaacatcatc
     1621 atgattttca tgaaattgtg attgtcgaac atggcacggg tattcatgtg tttaatgggc
     1681 agccctatac catcaccggt ggcacggtct gtttcgtacg cgatcatgat cggcatctgt
     1741 atgaacatac cgataatctg tgtctgacca atgtgctgta tcgctcgccg gatcgatttc
     1801 agtttctcgc cgggctgaat cagttgctgc cacaagagct ggatgggcag tatccgtctc
     1861 actggcgcgt taaccacagc gtattgcagc aggtgcgaca gctggttgca cagatggaac
     1921 agcaggaagg ggaaaatgat ttaccctcga ccgccagtcg cgagatcttg tttatgcaat
     1981 tactgctctt gctgcgtaaa agcagtttgc aggagaacct ggaaaacagc gcatcacgtc
     2041 tcaacttgct tctggcctgg ctggaggacc attttgccga tgaggtgaat tgggatgccg
     2101 tggcggatca attttctctt tcactgcgta cgctacatcg gcagcttaag cagcaaacgg
     2161 gactgacgcc tcagcgatac ctgaaccgcc tgcgactgat gaaagcccga catctgctac
     2221 gccacagcga ggccagcgtt actgacatcg cctatcgctg tggattcagc gacagtaacc
     2281 acttttcgac gctttttcgc cgagagttta actggtcacc gcgtgatatt cgccagggac
     2341 gggatggctt tctgcaacgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg
     2401 tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg
     2461 acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac
     2521 cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg
     2581 accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac
     2641 gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag
     2701 gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata
     2761 tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata
     2821 aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg
     2881 tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc
     2941 cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg
     3001 atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac
     3061 tgtacaaagg ctccggctcc ggctcccacc atcaccatca ccattgataa ctcgagcccc
     3121 aagggcgaca ccccataatt agcccgggcg aaaggcccag tctttcgact gagcctttcg
     3181 ttttatttga tgcctggcag ttccctactc tcgcatgggg agtccccaca ctaccatcgg
     3241 cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc gcgctactgc
     3301 cgccaggcaa acaaggggtg ttatgagcca tattcaggta taaatgggct cgcgataatg
     3361 ttcagaattg gttaattggt tgtaacactg acccctattt gtttattttt ctaaatacat
     3421 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3481 aggaagaata tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3541 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3601 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3661 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3721 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3781 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3841 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3901 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3961 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4021 accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4081 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4141 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatccgg agccggtgag
     4201 cgtggttctc gcggtatcat cgcagcgctg gggccagatg gtaagccctc ccgtatcgta
     4261 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4321 ataggtgcct cactgattaa gcattggtaa gcggcgcgcc atcgaatggc gcaaaacctt
     4381 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atatgaaacc
     4441 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     4501 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     4561 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     4621 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     4681 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     4741 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     4801 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     4861 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     4921 tgaggacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     4981 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     5041 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     5101 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     5161 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     5221 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagatagct catgttatat
     5281 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     5341 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc cagtctcact
     5401 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5461 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgactcat
     5521 gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc agaccccgta
     5581 gaaa
//
