LOCUS       pKG2197_pPV2-ZF1        3915 bp    DNA     circular SYN 30-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3915)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3915
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      49..637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     538..557
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     638..655
                     /label=oKL179
     primer_bind     640..680
                     /label=oKL276
     primer_bind     complement(640..657)
                     /label=pPV-bb-rev-v2
     primer_bind     complement(643..657)
                     /label=pPV1_bb_rev
     misc_feature    663..668
                     /locus_tag="Kozak(1)"
                     /label=Kozak
                     /label=Kozak(1)
     misc_feature    669..671
                     /locus_tag="Start(1)"
                     /label=Start
                     /label=Start(1)
     misc_feature    672..1199
                     /locus_tag="ZF10-1"
                     /label=ZF10-1
     misc_signal     1209..1325
                     /locus_tag="3xFlag-NLS"
                     /label=3xFlag-NLS
     misc_feature    1332..1349
                     /locus_tag="GS linker"
                     /label=GS linker
     misc_signal     1356..1385
                     /locus_tag="NS5A/5B cutsite (CC)"
                     /label=NS5A/5B cutsite (CC)
     misc_feature    1374..1397
                     /locus_tag="N-end rule"
                     /label=N-end rule
     misc_feature    1407..1424
                     /locus_tag="AU1"
                     /label=AU1
     exon            1440..1478
                     /locus_tag="NS4A"
                     /label=NS4A
     exon            1491..2057
                     /locus_tag="NS3"
                     /label=NS3
     misc_feature    1659..1661
                     /locus_tag="HIS-57, catalytic triad"
                     /label=HIS-57, catalytic triad
     misc_feature    1731..1733
                     /locus_tag="Asp 81, catalytic triad"
                     /label=Asp 81, catalytic triad
     misc_feature    1905..1907
                     /locus_tag="Ser 139, catalytic triad"
                     /label=Ser 139, catalytic triad
     misc_feature    1992..1994
                     /locus_tag="Asp168"
                     /label=Asp168
     misc_signal     2109..2150
                     /locus_tag="NS4A/4B cutsite (CS)"
                     /label=NS4A/4B cutsite (CS)
     misc_feature    2178..2195
                     /locus_tag="GS linker(1)"
                     /label=GS linker
                     /label=GS linker(1)
     misc_feature    2202..2774
                     /locus_tag="p65(aa361-551)"
                     /label=p65(aa361-551)
     primer_bind     complement(2761..2808)
                     /label=oKL277
     misc_feature    2781..2783
                     /locus_tag="Stop(1)"
                     /label=Stop
                     /label=Stop(1)
     primer_bind     2788..2808
                     /label=pPV-bb-fwd-v2
     primer_bind     2788..2802
                     /label=pPV1_bb_fwd
     primer_bind     complement(2789..2808)
                     /label=oKL247
     promoter        2828..2932
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2865..2886)
                     /label=oKL157
     primer_bind     complement(2865..2886)
                     /label=oSEQ095 oKL157
     CDS             join(2933..3001,3002..3793)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGYRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2933..2957
                     /label=oSEQ083 Amp-fwd
     primer_bind     complement(3151..3170)
                     /label=oSEQ066 Amp_rev
     primer_bind     complement(3347..3364)
                     /label=oKL048
     primer_bind     3347..3364
                     /label=oKL191
     primer_bind     complement(3354..3371)
                     /label=oKL070
ORIGIN
        1 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
       61 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg ggtctcaaat
      661 gagccaccat gtctagaccc ggagagcgcc cattccagtg tcggatttgc atgcggaact
      721 tttcgagaag acacggcctg gacagacata cccgtactca tacaggtgaa aaaccctttc
      781 agtgtcggat ctgtatgcga aatttctccg accacagcag cctgaagaga catctacgta
      841 cccacaccgg cagccagaag ccatttcagt gtcggatctg tatgcggaac ttctccgtga
      901 gacacaacct gaccagacat ctacgtacgc acaccggaga gaagccattc caatgccgaa
      961 tatgcatgcg caacttcagt gaccacagca acctgagcag acacctaaaa acccacaccg
     1021 gttcccagaa gccatttcag tgtcggatct gtatgcggaa cttctcccag cgcagcagcc
     1081 tggtgagaca tctacgtacg cacaccggag agaagccatt ccaatgccga atatgcatgc
     1141 gcaacttcag tgagagcggc cacctgaaga gacacctgcg tacgcacctg aggggatcca
     1201 cctgcaggga ctacaaagac catgacggtg attataaaga tcatgacatc gattacaagg
     1261 atgacgatga caagatggcc cccaagaaaa agaggaaggt gggcattcac ggggtgccgg
     1321 gtggactcga gggaggcggt ggaagcggcg gtaccgagga cgtggtgtgc tgccactcaa
     1381 tctacggcaa gaagaagggt gatatcgaca cctaccgata cataggctct tccgggacag
     1441 gctgcgtggt catagtgggc aggatcgtct tgtccggatc cggcactagt gcgcccatca
     1501 cggcgtacgc ccagcagacg agaggcctcc tagggtgtat aatcaccagc ctgactggcc
     1561 gggacaaaaa ccaagtggag ggtgaggtcc agatcgtgtc aactgctacc caaaccttcc
     1621 tggcaacgtg catcaatggg gtatgctggg cagtctacca cggggccgga acgaggacca
     1681 tcgcatcacc caagggtcct gtcatccaga tgtataccaa tgtggaccaa gaccttgtgg
     1741 gctggcccgc tcctcaaggt tcccgctcat tgacaccctg tacctgcggc tcctcggacc
     1801 tttacctggt cacgaggcac gccgatgtca ttcccgtgcg ccggcgaggt gatagcaggg
     1861 gtagcctgct ttcgccccgg cccatttcct acttgaaagg ctcctcgggg ggtccgctgt
     1921 tgtgccccgc gggacacgcc gtgggcctat tcagggccgc ggtgtgcacc cgtggagtgg
     1981 ctaaagcggt ggactttatc cctgtggaga acctagagac aaccatgaga tccccggtgt
     2041 tcacggacaa ctcctctcca ccagcagtca ccctgacgca cccaatcacc aaaatcgata
     2101 gggaggttct ctaccaggag ttcgatgaga tggaagagtg ctctcagcac tatccctacg
     2161 atgtgcccga ttacgctgga ggcggtggaa gcggcggtac cgatgagttt cccaccatgg
     2221 tgtttccttc tgggcagatc agccaggcct cggccttggc cccggcccct ccccaagtcc
     2281 tgccccaggc tccagcccct gcccctgctc cagccatggt atcagctctg gcccaggccc
     2341 cagcccctgt cccagtccta gccccaggcc ctcctcaggc tgtggcccca cctgccccca
     2401 agcccaccca ggctggggaa ggaacgctgt cagaggccct gctgcagctg cagtttgatg
     2461 atgaagacct gggggccttg cttggcaaca gcacagaccc agctgtgttc acagacctgg
     2521 catccgtcga caactccgag tttcagcagc tgctgaacca gggcatacct gtggcccccc
     2581 acacaactga gcccatgctg atggagtacc ctgaggctat aactcgccta gtgacagggg
     2641 cccagaggcc ccccgaccca gctcctgctc cactgggggc cccggggctc cccaatggcc
     2701 tcctttcagg agatgaagac ttctcctcca ttgcggacat ggacttctca gccctgctga
     2761 gtcagatcag ctcccaattg taacaacaga gacctagacg tcaggtggca cttttcgggg
     2821 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2941 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3001 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3061 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3121 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3241 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3481 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3541 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt atcgcggtat
     3661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3781 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3841 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3901 cccttaacgt gagtt
//
