LOCUS       40924_33327        7904 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNK207A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7904)
  AUTHORS   Kandul NP, Guo M, Hay BA.
  TITLE     A positive readout single transcript reporter for site-specific mRNA
            cleavage
  JOURNAL   PeerJ (2017) In press
REFERENCE   2  (bases 1 to 7904)
  AUTHORS   Kandul NP, Guo M, Hay BA.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JAN-2017) Biology and Biological Engineering, 
            California Institute of Technology, 1200 E. California Blvd., 
            Pasadena, CA 91125, USA
REFERENCE   3  (bases 1 to 7904)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7904)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PeerJ 
            (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-JAN-2017) Biology and Biological Engineering, California 
            Institute of Technology, 1200 E. California Blvd., Pasadena, CA 
            91125, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..460
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     601..617
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        627..645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        699..3208
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     misc_RNA        3239..3336
                     /product="CopT"
     CDS             3361..5007
                     /label=luciferase
                     /note="firefly luciferase"
     misc_RNA        5285..5374
                     /product="CopA"
     promoter        complement(5716..5734)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5755..5771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5779..5795)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5803..5833)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5848..5869)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6157..6745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6919..7776)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7777..7881)
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa atgagctgat ttaacaaaaa
      421 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
      481 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      541 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      601 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggccacgc
      661 tgaggaaggg ccggccttct taattaacta aaaaaaatca tgaatggcat caactctgaa
      721 tcaaatcttt gcagatgcac ctacttctca tttccactgt cacatcattt ttccagatct
      781 cgctgcctgt tatgtggccc acaaaccaag acacgtttta tggccattaa agctggctga
      841 tcgtcgccaa acaccaaata cataatgaat atgtacacat tcgagaaaga agcgatcaaa
      901 gaagcgtctt cgggcggagt aggagaatgc ggaggagaag gagaacgagc tgatctagta
      961 tctctccaca atccaatgcc aactgaccaa ctggccatat tcggagcaat ttgaagccaa
     1021 tttccatcgc ctggcgatcg ctccattctt ggctatatgt ttttcaccgt tacccggggc
     1081 cattttcaaa gactcgtcgg caagataaga ttgtgtcact cgctgtctct cttcatttgt
     1141 cgaagaatgc tgaggaattt cgcgatgacg tcggcgagta ttttgaagaa tgagaataat
     1201 ttgtatttat acgaaaatca gttagtggaa ttttctacaa aaacatgtta tctatagata
     1261 attttgttgc aaaatatgtt gactatgaca aagattgtat gtatatacct ttaatgtatt
     1321 ctcattttct tatgtattta taatggcaat gatgatactg atgatatttt aagatgatgc
     1381 cagaccaaaa ggcttgaatt tctgcgtctt ttgccgaacg cagtgcatgt gcaattgttg
     1441 ttttttggaa tattcaattt tcggactgtc cgctttgatt tcagtttctt ggcttattca
     1501 aaaagcaaag taaagccaaa aaagcgagat ggcaatacca aatgcggcaa aacggtagtg
     1561 gaaggaaagg ggtgcggggc agcggaagga agggtggggc ggggcgtggc ggggtctgtg
     1621 gctgggcgcg acgtcaccga cgttggagcc actcctttga ccatgtgtgc gtgtgtgtat
     1681 tattcgtgtc tcgccactcg ccggttgttt ttttcttttt atgctgcgct ctctctagcg
     1741 ccatctcgct tacgcatgct caacgcaccg catgttgccg tttcctttta tgcgtcattt
     1801 tggctcgaaa taggcaatta tttaaacaaa gattagtcaa cgaaaacgct aaaataaata
     1861 agtctacaat atggttactt attgccatgt gtgtgcagcc aacgatagca acaaaagcaa
     1921 caacacaggt ggctttccct ctttcacttt ttgtttgcaa gccgcgtgcg agcaagacgg
     1981 cacgaccggc aaacgcaatt acgctgacaa agagcagacg aagttttggc gaaaaacatc
     2041 aaggcgcctg atacgaatgc atttgcaata acaattgcga tatttaatat tgtttatgaa
     2101 gctgtttgac ttcaaaacac acaaaaaaaa aaataaaaca aattatttga aagagaatta
     2161 ggaatcggac gcttatcgtt agggtaacaa caagaaatgc ttactgagtc acagcctctg
     2221 gaaaactgcc gcaagccaga gagagagaga aaaagaggga gagcagctta gaccgcatgt
     2281 gcttgtgtgt gaggcgtctc tctcttcgtc tctgttgcgc aaacgcatag actgcactga
     2341 aaaaatcgat tacctatttt ttatgaatga atatttgcac tattactatt caaaactatt
     2401 aagatagcaa tcacattcaa tagccaaata ctataccacc tgagcgatgc aacgaaatga
     2461 tcaatttgag caaaaatgct gcatatttag gacggcatca ttatagaaat gcttcttgct
     2521 gtgtactttt ctctcgtctg gcagctgttt cgccgttatt gttaaaaccg gcttaagtta
     2581 ggtgtgtttt ctacgactag tgaatgccct actagaagat gtgtgttgca caaaatgtcc
     2641 ctggaataac caatttgaag tgcagatagc agtaaacgta agctaatatg aatattattt
     2701 aactgtaatg ttttaatatc gctggacatt actaataaac ccactataaa cacatgtaca
     2761 tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa tgtgtaaaag caccgtgacc
     2821 atcacagcat aaagataacc agctgaagta tcgaatatga gtaaccccca aattgaatca
     2881 catgccgcaa ctgataggac ccatggaagt acactcttca tggcgatata caagacacac
     2941 acaagcacga acacccagtt gcggaggaaa ttctccgtaa atgaaaaccc aatcggcgaa
     3001 caattcatac ccatatatgg taaaagtttt gaacgcgact tgagagcgga gagcattgcg
     3061 gctgataagg ttttagcgct aagcgggctt tataaaacgg gctgcgggac cagttttcat
     3121 atcactaccg tttgagttct tgtgctgtgt ggatactcct cccgacacaa agccgctcca
     3181 tcagccagca gtcgtctaat ccagagaccc cggatcgggc tagcagtagg gtaccaaaag
     3241 caaaaacccc gataatcttc ttcaactttg gcgagtacga aaagattacc ggggcccact
     3301 taaaccgtat agccaacaat tcagctatgc ggggagtata agtaagctta caaaatggct
     3361 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
     3421 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     3481 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
     3541 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     3601 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     3661 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
     3721 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     3781 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
     3841 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     3901 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     3961 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     4021 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     4081 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     4141 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     4201 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     4261 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     4321 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     4381 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     4441 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     4501 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     4561 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     4621 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     4681 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     4741 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     4801 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     4861 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     4921 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     4981 gccaagaagg gcggaaagat cgccgtgtaa gaattcgagg cggatccaaa aaaaaaaaaa
     5041 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5101 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5161 aaaaaaagcg gccgccactc ctcgagttcg aacaatagat actaccactc ctcgagttcg
     5221 aacaatcgta tcagtcccac tcctcgagtt cgaacaatcc gcggatcgac tagtgccgaa
     5281 ttcgtagctg aattgttggc tatacggttt aagtgggccc cggtaatctt ttcgtactcg
     5341 ccaaagttga agaagattat cggggttttt gcttctatct agactagagg cctgcgtcac
     5401 gccacttcaa cgctcgatgg gagcgtcatt ggtgggcggg gtaaccgtcg aaatcagtgt
     5461 ttacgcttcc aatcgcaaca aaaaattcac tgcaacactg aaaagcatac gaaaacgatg
     5521 aagattgtac gagaaaccat aaagtatttt atccacaaag acacgtatag cagaaaagcc
     5581 aagttaactc ggcgataagt tgtgtacaca agaataaaat cggccagatt cagtgttgtc
     5641 agaaataaga aaaccccact gtttttcttt ctgcaggata ccggtataac gcgaaacagt
     5701 ggccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     5761 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5821 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5881 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6661 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6721 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6781 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6841 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6901 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6961 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7021 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7081 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7141 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7201 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7261 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7321 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7381 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7441 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7501 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7561 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7621 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7681 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7741 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7801 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7861 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
