LOCUS       Exported                5820 bp DNA     circular SYN 20-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Jan 20, 2022 from SnapGene 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5820
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          878..1353
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     primer_bind     1375..1414
                     /label=pMD2_clone_f
     CDS             1428..2963
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSVGmut
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPQSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTEAELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     primer_bind     complement(1473..1514)
                     /label=VSVG_H8A_r
     primer_bind     1483..1527
                     /label=VSVG_H8A_f
     CDS             1499
                     /label=H8
     primer_bind     1597..1638
                     /label=VSVG_K47Q_f
     primer_bind     complement(1598..1642)
                     /label=VSVG_K47Q_r
     CDS             1615..1616
                     /label=K47Q
     primer_bind     complement(2081..2121)
                     /label=VSVG_Y209A_r
     primer_bind     2093..2130
                     /label=VSVG_Y209A_f
     CDS             2100..2102
                     /label=Y209
     primer_bind     complement(2509..2550)
                     /label=VSVG_R354A_r2
     primer_bind     2515..2556
                     /label=VSVG_R354A_f
     CDS             2535..2537
                     /label=R354A
     primer_bind     complement(3088..3127)
                     /label=pMD2_clone_r
     polyA_site      3163..3556
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      complement(4066..4654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4825..5685)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5686..5790)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatcccctg agggggcccc catgggctag aggatccggc ctcggcctct gcataaataa
       61 aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata tcataatatg
      121 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      781 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc ctcgaagctt
      841 acatgtggta ccgagctcgg atcctgagaa cttcagggtg agtctatggg acccttgatg
      901 ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag
      961 ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct
     1021 tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc
     1081 agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat
     1141 aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg
     1201 taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct
     1261 tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta
     1321 atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg
     1381 ctggcccatc actttggcaa agcacgtgag atctgaattc tgacactatg aagtgccttt
     1441 tgtacttagc ctttttattc attggggtga attgcaagtt caccatagtt tttccacaca
     1501 accaaaaagg aaactggaaa aatgttcctt ctaattacca ttattgcccg tcaagctcag
     1561 atttaaattg gcataatgac ttaataggca cagccttaca agtcaaaatg ccccagagtc
     1621 acaaggctat tcaagcagac ggttggatgt gtcatgcttc caaatgggtc actacttgtg
     1681 atttccgctg gtatggaccg aagtatataa cacattccat ccgatccttc actccatctg
     1741 tagaacaatg caaggaaagc attgaacaaa cgaaacaagg aacttggctg aatccaggct
     1801 tccctcctca aagttgtgga tatgcaactg tgacggatgc cgaagcagtg attgtccagg
     1861 tgactcctca ccatgtgctg gttgatgaat acacaggaga atgggttgat tcacagttca
     1921 tcaacggaaa atgcagcaat tacatatgcc ccactgtcca taactctaca acctggcatt
     1981 ctgactataa ggtcaaaggg ctatgtgatt ctaacctcat ttccatggac atcaccttct
     2041 tctcagagga cggagagcta tcatccctgg gaaaggaggg cacagggttc agaagtaact
     2101 actttgctta tgaaactgga ggcaaggcct gcaaaatgca atactgcaag cattggggag
     2161 tcagactccc atcaggtgtc tggttcgaga tggctgataa ggatctcttt gctgcagcca
     2221 gattccctga atgcccagaa gggtcaagta tctctgctcc atctcagacc tcagtggatg
     2281 taagtctaat tcaggacgtt gagaggatct tggattattc cctctgccaa gaaacctgga
     2341 gcaaaatcag agcgggtctt ccaatctctc cagtggatct cagctatctt gctcctaaaa
     2401 acccaggaac cggtcctgct ttcaccataa tcaatggtac cctaaaatac tttgagacca
     2461 gatacatcag agtcgatatt gctgctccaa tcctctcaag aatggtcgga atgatcagtg
     2521 gaactaccac agaagccgaa ctgtgggatg actgggcacc atatgaagac gtggaaattg
     2581 gacccaatgg agttctgagg accagttcag gatataagtt tcctttatac atgattggac
     2641 atggtatgtt ggactccgat cttcatctta gctcaaaggc tcaggtgttc gaacatcctc
     2701 acattcaaga cgctgcttcg caacttcctg atgatgagag tttatttttt ggtgatactg
     2761 ggctatccaa aaatccaatc gagcttgtag aaggttggtt cagtagttgg aaaagctcta
     2821 ttgcctcttt tttctttatc atagggttaa tcattggact attcttggtt ctccgagttg
     2881 gtatccatct ttgcattaaa ttaaagcaca ccaagaaaag acagatttat acagacatag
     2941 agatgaaccg acttggaaag taactcaaat cctgcacaac agattcttca tgtttggacc
     3001 aaatcaactt gtgataccat gctcaaagag gcctcaatta tatttgagtt tttaattttt
     3061 atgaaaaaaa aaaaaaaaac ggaattcacc ccaccagtgc aggctgccta tcagaaagtg
     3121 gtggctggtg tggctaatgc cctggcccac aagtttcact aagctcgctt ccttgctgtc
     3181 caatttctat taaaggttcc ttggttccct aagtccaact actaaactgg gggatattat
     3241 gaagggcctt gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat
     3301 gtatttaaat tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta
     3361 aaacataaag aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc
     3421 catgaaagaa ggtgaggctg caaacagcta atgcacattg gcaacagccc tgatgcctat
     3481 gccttattca tccctcagaa aaggattcaa gtagaggctt gatttggagg ttaaagtttg
     3541 gctatgctgt attttacatt acttattgtt ttagctgtcc tcatgaatgt cttttcacta
     3601 cccatttgct tatcctgcat ctctcagcct tgactccact cagttctctt gcttagagat
     3661 accacctttc ccctgaagtg ttccttccat gttttacggc gagatggttt ctcctcgcct
     3721 ggccactcag ccttagttgt ctctgttgtc ttatagaggt ctacttgaag aaggaaaaac
     3781 agggggcatg gtttgactgt cctgtgagcc cttcttccct gcctccccca ctcacagtga
     3841 cccggaatcc ctcgacatgg cagtctagca ctagtgcggc cgcagatctg cttcctcgct
     3901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4261 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4921 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5041 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5101 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5701 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5761 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
//
