LOCUS       40924_46158        6628 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pW1336, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  AUTHORS   Barrett SP, Wang PL, Salzman J.
  TITLE     Circular RNA biogenesis can proceed through an exon-containing 
            lariat precursor
  JOURNAL   Elife 4 (2015) In press
  PUBMED    26057830
REFERENCE   2  (bases 1 to 6628)
  AUTHORS   Wang PL.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2015) Biochemistry, Stanford University, 279 
            Campus Drive, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 6628)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6628)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife 4 
            (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUN-2015) Biochemistry, Stanford University, 279 Campus Drive, 
            Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(147..930)
                     /direction=LEFT
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(1280..1296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1509..1964
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2246..2350
                     /label=AmpR promoter
     CDS             2351..3208
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3382..3970
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4258..4279
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4294..4324
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4332..4348
                     /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)."
     primer_bind     4356..4372
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4876..5967)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5968..6375)
                     /label=LEU2 promoter
ORIGIN
        1 ttactagtat ggcccagccg gccatggttg gcgcgccact taattaactg cggccgcaac
       61 tcgagatgag ctcgaattcg agtctaactc cttaaccact cggacatagt gacttatctg
      121 acactcattg taaattaata tatatatagg cattttgttt agttaaaggt acttaagtaa
      181 ttagtataaa cgaaccaatt ttataatcag gaagttaagt gaatggtagc acatgtcgta
      241 aaaattgtga atttttattg aataatattt taaatacaag cctttctaga ctaggtatac
      301 tcataaacat atatgagcaa aaggatagag gagattacat tgcatcttct acaaatttat
      361 ttattgccct ttactgaaaa attaaataat gagtactaaa tgataaaaag cgctcagtac
      421 aagaaagatt gcaaaaatat tgcattcttc atgaattaaa gttgcatata aggcatattg
      481 aaagtaatag tactaaaaca gcagttagcg aaaattaata gaattatatt cgcaagacaa
      541 ttgtgacatt aaattaaaaa attgtagaat ttttactatc ctctttaacg ccatgagcct
      601 ttataaaaag gttaaattag ttttaacatt ctttttttga gtaagagttt acaatttatc
      661 aaaacctgtg ttattatatt cattagtttc aatttattag catctagaga aaaatcaatt
      721 ggcagttaca ttgttggaat ttatgaagaa aaagaatcta caacggagaa taagttgctg
      781 atcgctttcc ctaaaattgt atattttgct gagcttattt tgacatttcg ttgaagtttt
      841 ctctaattcg cattcatttt aagtaaaaca atgagaaata aaattacaaa aaatacaaat
      901 taaaatacaa tttttagcta taattataga cgatgccctt gtatcccatt ctgtctcgct
      961 tgcccctact ttttatcttt tatataccat aatgaacgct gccgctacta accatacccc
     1021 gattttacat ttcggactcc caaggacgta caaaatagaa aactatagaa aaaaataatc
     1081 agaaaatagc atgtcatctc tttgtaaaac gcgtttgcaa gaagaaagga aacaatggag
     1141 aagagatcat ccatttgtat gtaaatttta gtaaacttga agaaatcact aacaacttct
     1201 cttacttagg gattctatgc aaaaccttgt aaatcatctg atggaggact cgatttaatg
     1261 aattggaagg ttggaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1321 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1381 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     1441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     1501 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1561 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     1621 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1681 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     1741 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1801 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     1861 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     1921 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2041 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2101 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2161 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2221 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2281 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2341 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2401 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2461 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2521 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2581 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2641 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2701 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2761 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2821 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2881 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2941 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3001 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3061 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3121 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3181 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3241 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3301 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3361 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3421 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3481 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3541 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3601 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3661 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3721 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3781 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3841 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3901 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3961 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4021 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4081 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4141 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4201 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4261 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4321 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4381 cgccaagctt gtcgatcgac tacgtcgtta aggccgtttc tgacagagta aaattcttga
     4441 gggaactttc accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat
     4501 ggtcaggtca ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc
     4561 aatatataaa ttaggaatca tagtttcatg attttctgtt acacctaact ttttgtgtgg
     4621 tgccctcctc cttgtcaata ttaatgttaa agtgcaattc tttttcctta tcacgttgag
     4681 ccattagtat caatttgctt acctgtattc ctttacatcc tcctttttct ccttcttgat
     4741 aaatgtatgt agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa
     4801 tttcgtgtcg tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa
     4861 aagagaatct ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc
     4921 ggtggtactg ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt
     4981 aactgcatct tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat
     5041 tgcagcagac aagatagtgg cgatagggtt gaccttattc tttggcaaat ctggagcaga
     5101 accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga
     5161 cgcagatggc aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc
     5221 accaaacatg ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat
     5281 catggcggca gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat
     5341 ggtttcctcc acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa
     5401 ggaccaaata ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat
     5461 tctttgcact tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc
     5521 ttcctttctc ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt
     5581 acctttagca aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt
     5641 acatggtctt aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc
     5701 aggtctaaca ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc
     5761 aaccttcttg gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc
     5821 agcaccacca attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat
     5881 agctttaaga accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa
     5941 aacgacgatc ttcttagggg cagacattag aatggtatat ccttgaaata tatatatata
     6001 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     6061 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     6121 ttagtcatga acgcttctct attctatatg aaaagccggt tccgcggctc tcacctttcc
     6181 tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac
     6241 aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt gttctcgtta
     6301 tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg
     6361 agtattccca cagttaactg cggtcaagat atttcttgaa tcaggcgcct tagaccgctc
     6421 ggccaaacaa ccaattactt gttgagaaat agagtataat tatcctataa atataacgtt
     6481 tttgaacaca catgaacaag gaagtacagg acaattgatt ttgaagagaa tgtggatttt
     6541 gatgtaattg ttgggattcc atttttaata aggcaataat attaggtatg tagatatact
     6601 agaagttctc ctcgacaagc ttgcatgc
//
