LOCUS       Exported                6264 bp ds-DNA     circular SYN 04-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, May 7, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6264
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    85..372
                     /label=CAG enhancer
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    807..1542
                     /label=B-actin promoter
     misc_feature    1735..1740
                     /label=Kozak
     CDS             1741..3276
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPQSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTEAELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     CDS             1927..1929
                     /label=K47Q
     misc_feature    2848..2850
                     /label=R354A
     misc_feature    3351..3407
                     /label=lac operator
     misc_feature    3351..3407
                     /label=beta-globin poly(A) sequence
     polyA_signal    3351..3406
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3767..3783
                     /label=M13 rev
     protein_bind    3791..3807
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    3815..3845
                     /label=lac promoter
     promoter        complement(3815..3845)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    3860..3881
                     /label=CAP binding site
     protein_bind    3860..3881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    3940..4135
                     /label=SV40 promoter
     promoter        3940..4135
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     misc_feature    3986..4121
                     /label=SV40 ori
     rep_origin      3986..4121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4141..4275
                     /label=SV40 poly(A)
     polyA_signal    4141..4275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4514..5102
                     /label=ori
     rep_origin      complement(4514..5102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5273..6133
                     /label=AMP
     CDS             complement(5273..6133)
                     /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"
     misc_feature    6134..6238
                     /label=AmpR promoter
     promoter        complement(6134..6238)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgccacc
     1741 atgaagtgcc ttttgtactt agccttttta ttcattgggg tgaattgcaa gttcaccata
     1801 gtttttccac acaaccaaaa aggaaactgg aaaaatgttc cttctaatta ccattattgc
     1861 ccgtcaagct cagatttaaa ttggcataat gacttaatag gcacagcctt acaagtcaaa
     1921 atgccccaaa gtcacaaggc tattcaagca gacggttgga tgtgtcatgc ttccaaatgg
     1981 gtcactactt gtgatttccg ctggtatgga ccgaagtata taacacattc catccgatcc
     2041 ttcactccat ctgtagaaca atgcaaggaa agcattgaac aaacgaaaca aggaacttgg
     2101 ctgaatccag gcttccctcc tcaaagttgt ggatatgcaa ctgtgacgga tgccgaagca
     2161 gtgattgtcc aggtgactcc tcaccatgtg ctggttgatg aatacacagg agaatgggtt
     2221 gattcacagt tcatcaacgg aaaatgcagc aattacatat gccccactgt ccataactct
     2281 acaacctggc attctgacta taaggtcaaa gggctatgtg attctaacct catttccatg
     2341 gacatcacct tcttctcaga ggacggagag ctatcatccc tgggaaagga gggcacaggg
     2401 ttcagaagta actactttgc ttatgaaact ggaggcaagg cctgcaaaat gcaatactgc
     2461 aagcattggg gagtcagact cccatcaggt gtctggttcg agatggctga taaggatctc
     2521 tttgctgcag ccagattccc tgaatgccca gaagggtcaa gtatctctgc tccatctcag
     2581 acctcagtgg atgtaagtct aattcaggac gttgagagga tcttggatta ttccctctgc
     2641 caagaaacct ggagcaaaat cagagcgggt cttccaatct ctccagtgga tctcagctat
     2701 cttgctccta aaaacccagg aaccggtcct gctttcacca taatcaatgg taccctaaaa
     2761 tactttgaga ccagatacat cagagtcgat attgctgctc caatcctctc aagaatggtc
     2821 ggaatgatca gtggaactac cacagaagct gaactgtggg atgactgggc accatatgaa
     2881 gacgtggaaa ttggacccaa tggagttctg aggaccagtt caggatataa gtttccttta
     2941 tacatgattg gacatggtat gttggactcc gatcttcatc ttagctcaaa ggctcaggtg
     3001 ttcgaacatc ctcacattca agacgctgct tcgcaacttc ctgatgatga gagtttattt
     3061 tttggtgata ctgggctatc caaaaatcca atcgagcttg tagaaggttg gttcagtagt
     3121 tggaaaagct ctattgcctc ttttttcttt atcatagggt taatcattgg actattcttg
     3181 gttctccgag ttggtatcca tctttgcatt aaattaaagc acaccaagaa aagacagatt
     3241 tatacagaca tagagatgaa ccgacttgga aagtaagcta gcagatcttt ttccctctgc
     3301 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3361 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3421 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3481 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     3541 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3601 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     3661 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     3721 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     3781 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3841 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3901 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     3961 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4021 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4081 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4141 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4201 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4261 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5161 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5221 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5281 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5341 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5401 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5461 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5521 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5581 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5641 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5701 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5761 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5821 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5881 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5941 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6001 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6061 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6121 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6181 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6241 catttccccg aaaagtgcca cctg
//
