LOCUS       Exported                3305 bp ds-DNA     circular SYN 20-APR-2018
DEFINITION  Cloning vector pUC19, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3305)
  AUTHORS   New England Biolabs.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-OCT-2007) Research Department, New England Biolabs, 
            240 County Road, Ipswich, MA 01938, USA
REFERENCE   2  (bases 1 to 3305)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 20, 2018 from SnapGene Viewer 4.1.8
            http://www.snapgene.com
COMMENT     See also GenBank accession L09137.
FEATURES             Location/Qualifiers
     source          1..3305
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     -35_signal      131..136
                     /label=bla promoter (counter-clockwise)
                     /label=bla promoter (counter-clockwise); TTCAAA
                     /note="bla promoter (counter-clockwise); TTCAAA"
     -10_signal      152..157
                     /label=bla promoter (counter-clockwise)
                     /label=bla promoter (counter-clockwise); GAGACA
                     /note="bla promoter (counter-clockwise); GAGACA"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     sig_peptide     201..269
                     /gene="bla"
                     /label=bla
                     /label=bla(1)
                     /note="required for secretion to the periplasm; cleaved off
                     to form the mature beta-lactamase protein."
     rep_origin      1232..1820
                     /label=pMB1 origin of replication (counter-clockwise) (RN
                     /note="pMB1 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     -35_signal      1232..1237
                     /label=RNAII promoter (counter-clockwise)
                     /label=RNAII promoter (counter-clockwise); TTGAGA
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     -10_signal      1253..1258
                     /label=RNAII promoter (counter-clockwise)
                     /label=RNAII promoter (counter-clockwise); CGTAAT
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     misc_RNA        1268..1820
                     /label=RNAII transcript (complementary strand)
                     /note="RNAII transcript (complementary strand)"
     misc_RNA        complement(1271..1378)
                     /label=RNAI transcript
                     /note="RNAI transcript"
     -10_signal      complement(1387..1392)
                     /label=RNAI promoter (clockwise)
                     /label=RNAI promoter (clockwise); GCTACA
                     /note="RNAI promoter (clockwise); GCTACA"
     -35_signal      complement(1409..1414)
                     /label=RNAI promoter (clockwise)
                     /label=RNAI promoter (clockwise); TTGAAG
                     /note="RNAI promoter (clockwise); TTGAAG"
     protein_bind    2112..2124
                     /label=CAP protein binding site
                     /bound_moiety="catabolite activator protein (CAP)"
                     /note="CAP protein binding site"
     -35_signal      2144..2149
                     /label=Plac promoter (drives lacZalpha; counter-clockwise
                     /label=Plac promoter (drives lacZalpha; 
                     counter-clockwise(1)
                     /note="Plac promoter (drives lacZalpha; counter-clockwise);
                     TTTACA"
     -10_signal      2168..2173
                     /label=Plac promoter (drives lacZalpha
                     /label=Plac promoter (drives lacZalpha; counter-clockwise
                     /note="Plac promoter (drives lacZalpha; counter-clockwise);
                     TATGTT"
     gene            2218..2270
                     /gene="lacZalpha"
                     /label=lacZalpha
                     /label=lacZalpha(2)
     CDS             2218..2270
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="beta-galactosidase alpha fragment"
                     /label=beta-galactosidase alpha fragment
                     /label=beta-galactosidase alpha fragment(1)
                     /translation="MTMITPSLHACRSTLED"
     misc_feature    complement(2235..2270)
                     /gene="lacZalpha"
                     /label=lacZalpha
                     /label=lacZalpha(3)
                     /note="multiple cloning site (EcoRI-HindIII)"
     misc_feature    2271..2890
                     /label=BFP ORF
     misc_feature    complement(2402..2424)
                     /label=potential guide site 3
     misc_feature    complement(2434..2456)
                     /label=potential guide site 2
     misc_feature    2435
                     /label=Silent C->G mutation
     misc_feature    complement(2467..2489)
                     /label=L2 protospacer-PAM
     misc_feature    2467..2471
                     /label=L2 PAM (mutated in HDR template)
     misc_feature    2468..2471
                     /label=BFP mutation
     misc_feature    2472..2489
                     /label=L2 Guide (Corrected)
     misc_feature    complement(2501..2523)
                     /label=potential guide site 1
     misc_feature    2891..2904
                     /label=BFP ORF
                     /label=BFP ORF(1)
     gene            2905..3160
                     /gene="lacZalpha"
                     /label=lacZalpha
     CDS             2905..3160
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="beta-galactosidase alpha fragment"
                     /label=beta-galactosidase alpha fragment
                     /translation="MFTGRRFTTS*LGKPWRYPT*SPCSTSPFRQLA**RRGPHRSPFP
                     TVAQPEWRMAPDAVFSPYASVRYFTPHMVHSQYNLL*CRI"
     misc_feature    complement(2905..2910)
                     /gene="lacZalpha"
                     /label=lacZalpha
                     /label=lacZalpha(1)
                     /note="multiple cloning site (EcoRI-HindIII)"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc atggtgagca
     2281 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2341 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2401 ccctgaagtt catctgcacc accggcaagc tgccggtgcc ctggcccacc ctcgtgacca
     2461 ccctgacgta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2521 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2581 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2641 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2701 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2761 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2821 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2881 cccagtccaa gctgagcaaa gaccgaattc actggccgtc gttttacaac gtcgtgactg
     2941 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3001 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3061 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3121 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3181 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3241 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3301 cgcga
//
