LOCUS       Exported                4564 bp ds-DNA     circular UNA 06-JAN-2020
DEFINITION  Restriction Cloning ligation product.
ACCESSION   urn.local...1q-8u0e8pd
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 7, 2020 from SnapGene 4.3.8.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4564
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          6..24
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source synthetic DNA construct
     source          join(4148..4564,1..4,1897..4147)
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
                     /label=source Cloning vector pUC19
     misc_feature    1..1900
                     /label=T7 pro-IRES-EGFP-bgh (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:e-8u0c3z9"
     misc_feature    1..138
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     gene            1..5
                     /label=BamHI
                     /note="/created_by=sophliu"
     misc_feature    1..4
                     /label=Ligation
                     /label=nonstandard type: ligation
     promoter        6..24
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      139..725
                     /note="/created_by=haiqi"
                     /note="/label=IRES"
     CDS             726..1445
                     /label=EGFP
                     /note="/created_by=haiqi"
     misc_feature    1896..1900
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    join(1897..4564,1..4)
                     /label=pUC19 (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:26-8u07xxy"
     CDS             complement(join(1897..1930,4293..4564,1..4))
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="beta-galactosidase alpha fragment"
                     /label=lacZalpha CDS
                     /translation="MPRVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA*MTMITPSLHAC"
     gene            complement(join(1897..1930,4293..4564,1..4))
                     /gene="lacZalpha"
                     /label=lacZalpha gene
     misc_feature    1897..1900
                     /label=Ligation
                     /label=nonstandard type: ligation
     -10_signal      complement(1975..1980)
                     /label=-10 signal
                     /note="Plac promoter (drives lacZalpha; counter-clockwise);
                     TATGTT"
     -35_signal      complement(1999..2004)
                     /label=-35 signal
                     /note="Plac promoter (drives lacZalpha; counter-clockwise);
                     TTTACA"
     protein_bind    complement(2024..2036)
                     /label=CAP protein binding site protein bind
                     /bound_moiety="catabolite activator protein (CAP)"
                     /note="CAP protein binding site"
     rep_origin      complement(2328..2916)
                     /direction=LEFT
                     /label=rep origin
                     /note="pMB1 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     misc_RNA        complement(2328..2880)
                     /label=RNA
                     /note="RNAII transcript (complementary strand)"
     -35_signal      2734..2739
                     /label=RNAI promoter (clockwise)
                     /label=RNAI promoter (clockwise); TTGAAG -35 signal
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      2756..2761
                     /label=RNAI promoter (clockwise)
                     /label=RNAI promoter (clockwise); GCTACA -10 signal
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        2770..2877
                     /label=RNAI transcript
                     /note="RNAI transcript"
     -10_signal      complement(2890..2895)
                     /label=-10 signal
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(2911..2916)
                     /label=-35 signal
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(3087..3947)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla CDS
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3087..3947)
                     /gene="bla"
                     /label=bla gene
     sig_peptide     complement(3879..3947)
                     /gene="bla"
                     /label=bla signal peptide
                     /note="required for secretion to the periplasm; cleaved off
                     to form the mature beta-lactamase protein."
     -10_signal      complement(3991..3996)
                     /label=-10 signal
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(4012..4017)
                     /label=-35 signal
                     /note="bla promoter (counter-clockwise); TTCAAA"
     misc_feature    join(4543..4564,1..4,1897..1913)
                     /gene="lacZalpha"
                     /label=lacZalpha misc
                     /label=nonstandard type: polylinker
                     /note="multiple cloning site (EcoRI-HindIII)"
ORIGIN
        1 gatcctaata cgactcacta tagggagacc caagctggct agcgtttaaa cttaagcttg
       61 gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc cagcacagtg
      121 gcggccgctc gaggttaacc cctctccctc ccccccccct aacgttactg gccgaagccg
      181 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
      241 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
      301 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
      361 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
      421 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
      481 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
      541 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
      601 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
      661 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
      721 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      781 tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca
      841 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      901 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      961 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1021 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1081 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1141 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1201 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1261 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1321 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1381 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1441 agtaatgaat taattaagaa ttatcaccgc ttctattcag ccagtaaggc ctgtctttaa
     1501 tggcctccgg catgagacac tttctagagt cgagtctaga gggcccgttt aaacccgctg
     1561 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gttttcctaa taaaatgagg
     1621 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1681 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     1741 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     1801 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1861 gcgccctagc gcccgctcct ttcgctttct tccctctgca ggcatgcaag cttggcgtaa
     1921 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1981 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2041 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2101 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2161 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3001 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3061 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3121 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3181 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3241 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3301 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3361 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3421 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3481 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3541 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3601 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3661 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3721 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3781 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3841 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3901 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3961 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4021 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4081 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4141 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4201 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4261 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4321 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4381 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     4441 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     4501 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga gctcggtacc
     4561 cggg
//
