LOCUS       Exported                6708 bp ds-DNA     circular SYN 22-MAY-2017
DEFINITION  pUC19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 3, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6708
                     /organism="pUC19"
                     /mol_type="other DNA"
                     /label=source:pUC19
     source          1239..3306
                     /organism="pUC19"
                     /mol_type="other DNA"
                     /label=source:pUC19
     promoter        142..171
                     /label=lac_promoter
     misc_feature    185..207
                     /label=M13_pUC_rev_primer
     promoter        206..224
                     /label=M13_reverse_primer promoter
                     /label=M13_reverse_primer
     misc_feature    239..1238
                     /label=LAH
     misc_feature    1239..1304
                     /label=3XFLAG
     misc_feature    1305..1338
                     /label=LoxP+2
     misc_feature    1339..1344
                     /label=HindIII
     misc_feature    complement(1345..1393)
                     /label=Synthetic poly(A)
     misc_feature    complement(1394..1993)
                     /label=Puro
     misc_feature    complement(2048..2316)
                     /label=SV40 promoter
     misc_feature    complement(2317..2322)
                     /label=BamHI
     misc_feature    2323..2356
                     /label=LoxP+2(2)
     misc_feature    2357..2358
                     /label=LoxP+2(1)
     misc_feature    2357..2358
                     /label=Extra Bases
     misc_feature    2359..2364
                     /label=KpnI
     misc_feature    2365..2367
                     /label=Halo START
     CDS             2368..3255
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(2368..2394)
                     /label=DY 140
     CDS             3268..3288
                     /codon_start=1
                     /label=TEV protease site
                     /translation="EDLYFQS"
     misc_feature    3307..4306
                     /label=RAH
     misc_feature    3351
                     /label=C<A
     misc_feature    3353..3372
                     /label=Suz12 sgRNA
     misc_feature    4309..4464
                     /label=lacZ_a
     promoter        complement(4312..4328)
                     /label=M13_forward20_primer promoter
                     /label=M13_forward20_primer
     misc_feature    complement(4321..4343)
                     /label=M13_pUC_fwd_primer
     misc_feature    4656..4678
                     /label=pGEX_3_primer
     promoter        4837..4865
                     /label=AmpR_promoter
     CDS             4907..5767
                     /codon_start=1
                     /label=ORF frame 3
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4907..5767
                     /gene="Ampicillin"
                     /label=Ampicillin
     rep_origin      5922..6541
                     /label=pBR322_origin
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttac
      241 tgaaatgggc ctatgtgtct atcgttcaaa gcaagacctg gccaggcgcg gtggctcacg
      301 cctgtaatcc cagcactttg ggaagccgag gcaggtggat cacgaggtca ggagatcgag
      361 accagcctgg ccaaaatggt gaaaccctgt ctctactaaa aacacaaaaa attagccagg
      421 cgtggtggcg agcgcctgta atcccagcta ctctggaggc tgaggcagga gaatcgcttc
      481 aacccgggag gcgcaggttg tagggagacg aagtcgcgcc actggcactc cagcctgggc
      541 gacagaggga ggctccgtct caaaaaaaaa aaaaagcaag actcgcaaaa caccggattc
      601 cttctcagct ttctgagctg aaaaggtata gatcctaaag aagagaagag tgtatgtaag
      661 tacttctcaa ccagtttacc tctctgacca ctcttacggt gtttgggggc aacaaactaa
      721 ggatctagac tcgctaaacc gctcgctggg tccttctctc cccacaatgc accggggcca
      781 gcaggaaggc cgctccgacc cctaattttc ccgcgaattc agttcaaaga ggcggccggg
      841 cccgcgatcg gcagcgcgca cgggccaacc aagccgcgcc tccgcgagaa gcgcctccgc
      901 gtgactgacg ggggaatccg cgggccaacg ggctgggcgg cccggcggtg cgcgctcccg
      961 ccggccaatc agagcgccgg gcgggggaag tgggcggagc gaggccaggg tagggtgagc
     1021 ggcctccgaa gcggagcggg gctctgagga gacacttttt ttttcctccc tccttccctc
     1081 ctctcctcct cccttccctt cccctctcct cccctctctc ctccttcccc cctcggtccg
     1141 ccggagcctg ctggggcgag cggttggtat tgcaggcgct tgctctccgg ggccgcccgg
     1201 cgggtagctg gcggggggag gaggcaggaa ccgcgatgga ctacaaagac catgacggtg
     1261 attataaaga tcatgatatc gattacaagg atgacgatga caagataact tcgtataatg
     1321 tatgctatac gaagttataa gcttcacaca aaaaaccaac acacagatgt aatgaaaata
     1381 aagatatttt atttcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     1441 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     1501 gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     1561 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca
     1621 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     1681 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
     1741 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
     1801 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
     1861 gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
     1921 cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
     1981 tgtactcggt catgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     2041 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     2101 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     2161 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     2221 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     2281 ttctgcctgc tggggagcct ggggactttc cacaccggat ccataacttc gtataatgta
     2341 tgctatacga agttatccgg taccatggca gaaatcggta ctggctttcc attcgacccc
     2401 cattatgtgg aagtcctggg cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc
     2461 acccctgtgc tgttcctgca cggtaacccg acctcctcct acgtgtggcg caacatcatc
     2521 ccgcatgttg caccgaccca tcgctgcatt gctccagacc tgatcggtat gggcaaatcc
     2581 gacaaaccag acctgggtta tttcttcgac gaccacgtcc gcttcatgga tgccttcatc
     2641 gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg actggggctc cgctctgggt
     2701 ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat ggagttcatc
     2761 cgccctatcc cgacctggga cgaatggcca gaatttgccc gcgagacctt ccaggccttc
     2821 cgcaccaccg acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg
     2881 ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg
     2941 ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc
     3001 ggtgagccag cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc
     3061 cctgtcccga agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc
     3121 gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat
     3181 ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtcgacg
     3241 ctcgagattt ccggcgagcc aaccactgag gatctgtact ttcagagcga taacgcgatc
     3301 ggtaccgcgc ctcagaagca cggcggtggg ggagggggcg gctcggggcc aagcgcgggg
     3361 tccgggggag gcggcttcgg gggttcggcg gcggtggcgg cggcgacggc ttcgggcggc
     3421 aaatccggcg gcgggagctg tggagggggt ggcagttact cggcctcctc ctcctcctcc
     3481 gcggcggcag cggcgggggc tgcggtgtta ccggtgaaga agccgaaaat ggagcacgtc
     3541 caggctgacc acgagctttt cctccaggcc tttgagagtg agtgtgtgcg aggctttgag
     3601 ggcaggaaga cccactctgc caacaccggg gatgggacac tctgctgggc ccccttcctc
     3661 ctcgggagtc cacttgtgtg gtagtggagg gaggaattga ggggatgtcc ccctttcgga
     3721 gattacgtct ccaggggatc cggagtccag ggccttggga gagctggaaa agagggtccg
     3781 ggcgggttct tgactccccc aagattgacg caagaggctc gggactgggg ccccacgttc
     3841 agagcaggcg gcttttagat gaggaggtag cagttgaact gagccgcagt cacatcccga
     3901 gactggttct tgttttcttt catgcctttc ggactctaga aatacagggg tttcggttgt
     3961 agcttttgag gggcaaaatg aagcctgtga agggatgagt gcagtggaac cgaagaaagt
     4021 gtccttagtc tgaaatgcct tcgcgctaac ttggtgctgc aggttcctat ggcacctttg
     4081 gatgcttaaa acccttcaaa tctatccacc cattttgtgg atatgtaaac tgagccagag
     4141 agtggtcaag catcttctct ggagacgacc ccggaattct gctttttcta cctggcttac
     4201 agaacatttt cttgcgtcaa tttctgtttc taagttctgt ctggaaggtg aaactgactt
     4261 atttggaaag gagagcaagg tgtctggaaa caaacaatag aatttaaatt cactggccgt
     4321 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     4381 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     4441 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     4501 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     4561 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4621 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4681 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4741 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4801 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4861 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4921 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4981 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5041 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5101 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5161 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5221 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5281 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5341 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5401 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5461 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5521 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5581 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5641 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5701 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5761 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5821 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5881 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5941 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6001 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6061 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6121 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6181 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6241 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6301 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6361 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6421 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6481 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6541 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6601 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6661 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
