LOCUS       New                     7756 bp    DNA     circular SYN 27-Jul-2017
DEFINITION  -
ACCESSION   -
KEYWORDS    -
SOURCE      -
FEATURES             Location/Qualifiers
     source          1..7756
                     /mol_type="other DNA"
                     /note="/dnas_title=""New Orco Plasmid"""
                     /organism="Cloning vector pUC19"
     gene            complement(146..416)
                     /gene="lacZalpha"
                     /note="/dnas_title=""lacZalpha"""
     CDS             complement(146..416)
                     /codon_start=1
                     /gene="lacZalpha"
                     /note="/dnas_title=""beta-galactosidase alpha fragment"""
                     /product="beta-galactosidase alpha fragment"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    417..1626
                     /label=left homology arm
     misc_feature    1594..1615
                     /label=Orco sgRNA 4
                     /note="modified to prevent recutting"
     misc_feature    1624..1626
                     /label=last codon of Orco
     misc_feature    1627..1683
                     /note="T2A"
     misc_feature    1684..2739
                     /note="QF2"
     misc_feature    2744..3000
                     /note="SV40 Term"
     misc_feature    3001..3246
                     /note="3xP3"
     misc_feature    3276..3953
                     /note="dsRed"
     misc_feature    3968..4195
                     /note="SV40"
     misc_feature    4196..5492
                     /label=right homology arm
     -10_signal      complement(5584..5589)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
     -35_signal      complement(5608..5613)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
     protein_bind    complement(5633..5645)
                     /bound_moiety="catabolite activator protein (CAP)"
                     /note="CAP protein binding site"
     rep_origin      complement(5937..6525)
                     /label=pMB1 ORI
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
     misc_RNA        complement(5937..6489)
                     /note="RNAII transcript (complementary strand)"
     -35_signal      6343..6348
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      6365..6370
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        6379..6486
                     /note="RNAI transcript"
     -10_signal      complement(6499..6504)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(6520..6525)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     gene            complement(6696..7556)
                     /gene="bla"
                     /note="/dnas_title=""bla"""
     CDS             complement(6696..7556)
                     /codon_start=1
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /note="/dnas_title=""beta-lactamase"""
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     sig_peptide     complement(7488..7556)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
                     /note="/dnas_title=""bla"""
     -10_signal      complement(7600..7605)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(7621..7626)
                     /note="bla promoter (counter-clockwise); TTCAAA"
BASE COUNT      2092 A      1842 C      1760 G      2062 T         0 OTHER
ORIGIN      ?
        1 TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA
       61 CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG
      121 TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC
      181 ACCATATGCG GTGTGAAATA CCGCACAGAT GCGTAAGGAG AAAATACCGC ATCAGGCGCC
      241 ATTCGCCATT CAGGCTGCGC AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT
      301 TACGCCAGCT GGCGAAAGGG GGATGTGCTG CAAGGCGATT AAGTTGGGTA ACGCCAGGGT
      361 TTTCCCAGTC ACGACGTTGT AAAACGACGG CCAGTGAATT CGAGCTCGGT ACCCGGGAGT
      421 TTCGCTTTTC CACGCGCGGT GTGTCTGGAA TCGTTATTAA CGAAAAAAAA TATTCTAATT
      481 CGGGATTTAC AATTCTCTGT GAATATGAAA ACATTTCAAC GAGAGAAACG AAAGTTATAG
      541 TGATTTGAAG GTTTTCGGCT ATTCTGTAAG ATATTCTTTC AAATAATAGG TGCCAAATTG
      601 GTGGACATGT TTTGTAATTA ACGATTTCAG ACACACCGTG CCACGTTTGG TTCTCTCTAG
      661 CAATTTGATT GTGCATTTTT TTTTCTCTAA CAGAGTTCAT CAGTGATGGA GGCTGCCTAC
      721 TCGTGCCACT GGTATGACGG TTCCGAGGAA GCCAAAACTT TCGTGCAAAT CGTTTGCCAG
      781 CAGTGTCAGA AAGCGATGAC CATATCCGGG GCCAAGTTTT TCACCGTTTC ACTGGATCTG
      841 TTCGCATCGG TGAGTGATTT TTTTTTCGTT TTCGATTCGG TTTCTGGGGG GAAATTAGTG
      901 TCGGATCGAT CGATAAAAGC GATTGCATGG GCATAACCAG AAAGGAGACA TGAGTCCTCC
      961 TGCCTCCACC TCCCTTTTGA AAAAAAGCTG GGTGAATTAA TTTTGTGTTT GTATTTTCAA
     1021 AATCGTCTAT ATCTACAAAA TAATTGCATC ACGATTGATA TTTCTGAGAA ATTTTTGAAG
     1081 TTACTCAAAC AATAATTTTC CTAAAAAAGA TCTTATCAGA AATTATGAGA AGGTTTTATC
     1141 AACAGTCCGC ATTTTTGAAT CAATATAATA AGTTTTTCTG AAAAAAAAAA AAAAAAATAC
     1201 AGAAATACGG TGACGAATTT CTGTGGAATT TTTTTGAGAT TCAAATATTT CAAAAACATA
     1261 TTTTATGGTC GTCTGCAAAA AACCGTGAGA CGTTTTTGAT TTGGTGGAAT CTATCTTGAA
     1321 GATTTCTTCG ATGAACTGTA TGAAATTACT ATGTCGAATC AATTGAATCT GTAGAAAAAA
     1381 TGTCTCATCC GGATTTTTCC AAGCCTTGTT TCTAATCGTC CACTGGACTC CACTTTGGAA
     1441 AAGTGTGAAT TTTTTAAGTC ACTTGCCCCT CCGGCTATTC TGGCTACGCC CATGACCGAT
     1501 TGTATAAGAA AGCCACGAAA CTGAGTCAAC TATAATTAAT GATTCGTCGT TTTCTTTCCG
     1561 TTTTCCTTTC CCCCACAGGT TCTGGGAGCG GTCGTCACCT ACTTCATGGT GCTCGTGCAG
     1621 TTGAAAGGAG AGGGCAGAGG AAGTCTTCTA ACATGCGGTG ACGTGGAGGA GAATCCCGGC
     1681 CCTATGCCAC CCAAGCGCAA AACGCTTAAC GCTGCGGCTG AGGCTAACGC TCATGCCGAC
     1741 GGACACGCCG ACGGAAACGC CGACGGACAC GTGGCCAATA CGGCCGCGTC CTCGAATAAT
     1801 GCGAGGTTCG CTGATCTCAC TAACATCGAT ACTCCGGGTC TGGGACCCAC AACTACGACC
     1861 CTGCTCGTGG AACCAGCACG CTCAAAGCGT CAACGAGTGT CCCGCGCATG CGACCAGTGC
     1921 CGTGCAGCCC GAGAGAAATG CGACGGAATA CAGCCTGCGT GTTTCCCGTG CGTTTCCCAG
     1981 GGAAGGTCCT GCACTTATCA GGCTTCGCCG AAAAAGAGGG GAGTTCAAAC CGGTTATATT
     2041 CGTACGCTGG AGCTCGCCCT CGCCTGGATG TTTGAAAATG TCGCGCGTTC CGAAGATGCC
     2101 TTGCATAACC TCCTCGTCCG TGACGCCGGA CAAGGATCAG CTCTGCTCGT TGGTAAAGAT
     2161 TCGCCGGCTG CCGAGCGACT CCATGCCCGT TGGGCTACTA GCCGTGTCAA TAAGAGCATT
     2221 ACCCGCCTCC TCCGTCAGTT GGAGCTCCCT CCTACCGCCA CGGCTACGGC CTCGATAATG
     2281 CCGCACGTGA TGGAGCAGCC TCTCAGTACC AGCATTAACC CCGTCAACGA CCGCTTCAAC
     2341 GGTATTCCCA ACCCCACTCC GTATAACTCC GATGCAGCTC TCGATGCTAT CACTCAGACC
     2401 AACGATTATG GAAGCGTAAA TACACATGGT ATCCTCTCTA CTTACCCGCC ACCGGCTACG
     2461 CACCTTAATG AAGCTTCCGT CGCTCTCGCT CCCGGTGGCG CCCCCCCCCG ACCGCCTCCT
     2521 CCGTATGTTG ACAGCACGAC CAATCACCCG CCGTACCACT CGAATCTGGT TCCAATGGCG
     2581 AACTTTGGTT ACTCGACCGT TGATTACGAT GCCATGGTTG ACGATTTGGC TAGCATTGAA
     2641 TACACGGACG CTGTGGATGT CGACCCACAG TTTATGACCA ATCTGGGATT CGTTCCTGGA
     2701 TGTAACTTCT CCGACATTAA TACATACGAA CAGTGATGAC GTCTATCGAT ACCGTCGACT
     2761 AAAGCCAAAT AGAAAATTAT TCAGTTCCTG GCTTAAGTTT TTAAAAGTGA TATTATTTAT
     2821 TTGGTTGTAA CCAACCAAAA GAATGTAAAT AACTAATACA TAATTATGTT AGTTTTAAGT
     2881 TAGCAACAAA TTGATTTTAG CTATATTAGC TACTTGGTTA ATAAATAGAA TATATTTATT
     2941 TAAAGATAAT TGCGTTTTTA TTGTCAGGGA GTGAGTTTGC TTAAAAACTC GTTTAGATCC
     3001 AATTCGAGCT CGCCCGGGGA TCTAATTCAA TTAGAGACTA ATTCAATTAG AGCTAATTCA
     3061 ATTAGGATCC AAGCTTATCG ATTTCGAACC CTCGACCGCC GGAGTATAAA TAGAGGCGCT
     3121 TCGTCTACGG AGCGACAATT CAATTCAAAC AAGCAAAGTG AACACGTCGC TAAGCGAAAG
     3181 CTAAGCAAAT AAACAAGCGC AGCTGAACAA GCTAAACAAT CGGGGTACCG CTAGAGTCGA
     3241 CGGTACCGCG GGCCCGGGAT CCACCGGTCG CCACCATGGC CTCCTCCGAG AACGTCATCA
     3301 CCGAGTTCAT GCGCTTCAAG GTGCGCATGG AGGGCACCGT GAACGGCCAC GAGTTCGAGA
     3361 TCGAGGGCGA GGGCGAGGGC CGCCCCTACG AGGGCCACAA CACCGTGAAG CTGAAGGTGA
     3421 CCAAGGGCGG CCCCCTGCCC TTCGCCTGGG ACATCCTGTC CCCCCAGTTC CAGTACGGCT
     3481 CCAAGGTGTA CGTGAAGCAC CCCGCCGACA TCCCCGACTA CAAGAAGCTG TCCTTCCCCG
     3541 AGGGCTTCAA GTGGGAGCGC GTGATGAACT TCGAGGACGG CGGCGTGGCG ACCGTGACCC
     3601 AGGACTCCTC CCTGCAGGAC GGCTGCTTCA TCTACAAGGT GAAGTTCATC GGCGTGAACT
     3661 TCCCCTCCGA CGGCCCCGTG ATGCAGAAGA AGACCATGGG CTGGGAGGCC TCCACCGAGC
     3721 GCCTGTACCC CCGCGACGGC GTGCTGAAGG GCGAGACCCA CAAGGCCCTG AAGCTGAAGG
     3781 ACGGCGGCCA CTACCTGGTG GAGTTCAAGT CCATCTACAT GGCCAAGAAG CCCGTGCAGC
     3841 TGCCCGGCTA CTACTACGTG GACGCCAAGC TGGACATCAC CTCCCACAAC GAGGACTACA
     3901 CCATCGTGGA GCAGTACGAG CGCACCGAGG GCCGCCACCA CCTGTTCCTG TAGCGGCCGC
     3961 GACTCTAGAT CATAATCAGC CATACCACAT TTGTAGAGGT TTTACTTGCT TTAAAAAACC
     4021 TCCCACACCT CCCCCTGAAC CTGAAACATA AAATGAATGC AATTGTTGTT GTTAACTTGT
     4081 TTATTGCAGC TTATAATGGT TACAAATAAA GCAATAGCAT CACAAATTTC ACAAATAAAG
     4141 CATTTTTTTC ACTGCATTCT AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTATAATT
     4201 CGTTAATTAT GGACAAGCTC AAGACTATCC GATACTATTT TTGATACTAA TGCAACTGAA
     4261 CGGCGATACA TGACATGACA TCGTTTTTAT ATGTAGATTT ATAAGGAAGA AAAAAATTAT
     4321 CAGTACAAGA ACAATTAGAC TAGACCCTCC GACACTGAGT AGCGGAGGCT AATGATGTTG
     4381 ATATATTAAA TATTTTAGTA AGAATTTTTG ATGTAGCTGA AATAGACGCT CAAGATGAAT
     4441 GCAATGAACT CTCGAGAACT TATTCATAAC AGTGAGAGAA TGCCCTGATT GTTCACTGTA
     4501 ATTAGTTTTC CACACGATTT ACTTGGTTTA TGAAAAGCCC TTGTAGATTG AGAAGTCATT
     4561 AGTCAAGATA TGTCACTCCC GTGCAAAAGT TTGGGTTCAC CCCCTCAAAC ATATACAAAA
     4621 GTGTTCTGTC CATATCTCTG TGATTACACG TCAAATTTAA ACTCTTTAAG CCGCATTCGA
     4681 AAGGCAAAGA GTTATTCTTA CTTCGTATGT ATTTTTCCAA AAACATTTTT TTAAATTTTG
     4741 TATACTAAAT TTTTATTAAA GTTGTGACAT TTTTCAAAAA ACACCCTTAA AAATCATATC
     4801 TAATTTCCTC AGCATTGGGT TGACCAAAAT TTCAAATCAA AGTGTCATTA GAATCGTAAT
     4861 CTTATATTCC TTGAAGAGAC ACCACGAAAT TTCGGCGGAC AAATCTGGAA AGTATTCAAA
     4921 ATCAATGAAA CAGTCAGTCA AGTCATCGTG CAAAAGTTTG GGTTCATCTC TCAGTATGAT
     4981 GCAAATCGTG CAAAAGTTTG GGTTCACCTG ATCCGCACGC ACATAATTTT TGTCAATATC
     5041 TCGGCCATTT TTCAACCGAT TTTAATAGTT TAAAGCTTTT TTGAGCGCAA ATAATGGCGT
     5101 GTACATGATT GGTCTGAGAT TTCACAGATT TAAGTAAGTT CAGGTGAACC CAAATTTTTG
     5161 CACGATACAC CATACACTGA GGGGTGAACC CAAACTTTTG CACGATGACT TGACTGACTG
     5221 TTTTATTGAT TTTGAATACT TCTCAGATTT TTCCGCCAAA ATTTAATGAG TGCTCTTCAA
     5281 AGTATATAAG ATTACGATTC TAATGACACC TTGTTTTAAA ATTTTGGTCA ATTGAATGCT
     5341 GAGGAAATTA GATATGATTT TTCAGTGTGT TTTTTGAAAA ATGTCCCAAC TTTAAGTACA
     5401 AATTTAGTAT ACAAAGTTCA AAGAATGTTT TTGGAAAAAT ACATACGAAG TAAGAATAAC
     5461 TCTTTGCCTT TCGAATGCGG CTTAGAGAGT TTTCTAGAGT CGACCTGCAG GCATGCAAGC
     5521 TTGGCGTAAT CATGGTCATA GCTGTTTCCT GTGTGAAATT GTTATCCGCT CACAATTCCA
     5581 CACAACATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG GTGCCTAATG AGTGAGCTAA
     5641 CTCACATTAA TTGCGTTGCG CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG
     5701 CTGCATTAAT GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC
     5761 GCTTCCTCGC TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT
     5821 CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG
     5881 TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC
     5941 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA
     6001 AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT
     6061 CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG
     6121 GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG
     6181 CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT
     6241 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC
     6301 AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC
     6361 TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC
     6421 GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT
     6481 TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC
     6541 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG
     6601 AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA
     6661 ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA
     6721 CCTATCTCAG CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG
     6781 ATAACTACGA TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC
     6841 CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC
     6901 AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT
     6961 AGAGTAAGTA GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC
     7021 GTGGTGTCAC GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG
     7081 CGAGTTACAT GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC
     7141 GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC ACTGCATAAT
     7201 TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA CTGGTGAGTA CTCAACCAAG
     7261 TCATTCTGAG AATAGTGTAT GCGGCGACCG AGTTGCTCTT GCCCGGCGTC AATACGGGAT
     7321 AATACCGCGC CACATAGCAG AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG
     7381 CGAAAACTCT CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA
     7441 CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC AAAAACAGGA
     7501 AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA AATGTTGAAT ACTCATACTC
     7561 TTCCTTTTTC AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA
     7621 TTTGAATGTA TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG
     7681 CCACCTGACG TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC
     7741 ACGAGGCCCT TTCGTC
//
