LOCUS       Exported                7681 bp ds-DNA     circular SYN 11-OCT-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Leonard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 11, 2021 from SnapGene 5.1.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7681
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate earlypromoter"
     promoter        645..663
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          714..1189
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     CDS             1251..2783
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     misc_feature    2785..2804
                     /label=ASSGSGS spacer
     CDS             2805..2831
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2885..3109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3155..3583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
     promoter        3597..3926
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3777..3912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3975..5000
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5130..5251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5300..5316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5324..5340
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5348..5378)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5393..5414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
     rep_origin      complement(5702..6287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6458..7318)
                     /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(7319..7423)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgtccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      661 agggagaccc aagcttacat gtggtaccga gctcggatcc tgagaacttc agggtgagtc
      721 tatgggaccc ttgatgtttt ctttcccctt cttttctatg gttaagttca tgtcatagga
      781 aggggagaag taacagggta cacatattga ccaaatcagg gtaattttgc atttgtaatt
      841 ttaaaaaatg ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc
      901 ctaatctctt tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct
      961 aaagaataac agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat
     1021 ttctgcatat aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc
     1081 cagctaccat tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag
     1141 ctaggccctt ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa
     1201 cgtgctggtc tgtgtgctgg cccatcactt tggcaaagaa ttctgacact atgaagtgcc
     1261 ttttgtactt agccttttta ttcattgggg tgaattgcaa gttcaccata gtttttccac
     1321 acaaccaaaa aggaaactgg aaaaatgttc cttctaatta ccattattgc ccgtcaagct
     1381 cagatttaaa ttggcataat gacttaatag gcacagcctt acaagtcaaa atgcccaaga
     1441 gtcacaaggc tattcaagca gacggttgga tgtgtcacgc ttccaaatgg gtcactactt
     1501 gtgatttccg ctggtatgga ccgaagtata taacacattc catccgatcc ttcactccat
     1561 ctgtagaaca atgcaaggaa agcattgaac aaacgaaaca aggaacttgg ctgaatccag
     1621 gcttccctcc tcaaagttgt ggatatgcaa ctgtgacgga tgccgaagca gtgattgtcc
     1681 aggtgactcc tcaccatgtg ctggttgatg aatacacagg agaatgggtt gattcacagt
     1741 tcatcaacgg aaaatgcagc aattacatat gccccactgt ccataactct acaacctggc
     1801 attctgacta taaggtcaaa gggctatgtg attctaacct catttccatg gacatcacct
     1861 tcttctcaga ggacggagag ctatcatccc tgggaaagga gggcacaggg ttcagaagta
     1921 actactttgc ttatgaaact ggaggcaagg cctgcaaaat gcaatactgc aagcattggg
     1981 gagtcagact cccatcaggt gtctggttcg agatggctga taaggatctc tttgctgcag
     2041 ccagattccc tgaatgccca gaagggtcaa gtatctctgc tccatctcag acctcagtgg
     2101 atgtaagtct aattcaggac gttgagagga tcttggatta ttccctctgc caagaaacct
     2161 ggagcaaaat cagagcgggt cttccaatct ctccagtgga tctcagctat cttgctccta
     2221 aaaacccagg aaccggtcct gctttcacca taatcaatgg taccctaaaa tactttgaga
     2281 ccagatacat cagagtcgat attgctgctc caatcctctc aagaatggtc ggaatgatca
     2341 gtggaactac cacagaaagg gaactgtggg atgactgggc accatatgaa gacgtggaaa
     2401 ttggacccaa tggagttctg aggaccagtt caggatataa gtttccttta tacatgattg
     2461 gacatggtat gttggactcc gatcttcatc ttagctcaaa ggctcaggtg ttcgaacatc
     2521 ctcacattca agacgctgct tcgcaacttc ctgatgatga gagtttattt tttggtgata
     2581 ctgggctatc caaaaatcca atcgagcttg tagaaggttg gttcagtagt tggaaaagct
     2641 ctattgcctc ttttttcttt atcatagggt taatcattgg actattcttg gttctccgag
     2701 ttggtatcca tctttgcatt aaattaaagc acaccaagaa aagacagatt tatacagaca
     2761 tagagatgaa ccgacttgga aaggctagca gtggctctgg tagctaccca tacgatgtgc
     2821 cagattacgc ttaagcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     2881 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2941 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3001 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3061 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3121 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3181 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3241 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3301 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3361 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3421 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3481 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3541 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3601 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3661 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3721 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3781 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3841 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3901 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3961 ctgatcagca cgtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     4021 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     4081 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     4141 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     4201 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     4261 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     4321 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     4381 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     4441 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     4501 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     4561 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     4621 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     4681 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     4741 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     4801 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     4861 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     4921 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     4981 gtccgagggc aaaggaatag cacgtgctac gagatttcga ttccaccgcc gccttctatg
     5041 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5101 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5161 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5221 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5281 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5341 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5401 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5461 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5521 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5581 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6121 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     6241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6301 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6361 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6421 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6481 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6541 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6601 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6661 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6721 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6781 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6841 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6901 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6961 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7021 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7081 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7141 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7201 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7261 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7321 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7381 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7441 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     7501 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     7561 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     7621 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     7681 t
//
