LOCUS       Exported                8467 bp ds-DNA     circular SYN 01-AUG-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8467)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Nov 22, 2020 from SnapGene 5.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8467
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1825..1942
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    2024..2248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2288..2308)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2312..2332)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2336..2356)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2369..3085)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     2379..2400
                     /label=seq egfp 27-4
     misc_feature    3101..3596
                     /label=PKGp
     primer_bind     complement(3113..3130)
                     /label=ATGCTCCAGACTGCCTTG
     CDS             3615..4316
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     misc_feature    4327..4915
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4798..4809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4988..5221
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5293..5414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5454..5589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5610..5628)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5638..5654)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5796..6251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6277..6381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6382..7242
                     /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"
     rep_origin      7413..8001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8289..8310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8325..8355
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8363..8379
                     /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)."
     primer_bind     8387..8403
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8424..8442
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgat
     1801 acaaatggca gtattcattc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     1861 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     1921 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggtc
     1981 tagtggatcc tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca
     2041 tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
     2101 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
     2161 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
     2221 acagatggct ggcaactaga aggcacagtc gaggctgatc agcttaatta aggatcctct
     2281 aggatcctac ctttctcttc ttttttggat ctacctttct cttctttttt ggatctacct
     2341 ttctcttctt ttttggatca gctcgagtct tgtacagctc gtccatgccg agagtgatcc
     2401 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2461 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2521 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     2581 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     2641 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     2701 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     2761 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     2821 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     2881 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     2941 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     3001 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     3061 acagctcctc gcccttgctc accatggtgg cggccaattc cagtactttt cccaaggcag
     3121 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     3181 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     3241 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     3301 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     3361 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     3421 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     3481 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     3541 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgaagc
     3601 ttgccgccgg caccatggtg agcaagggcg aggaggtcat caaagagttc atgcgcttca
     3661 aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
     3721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
     3781 ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc
     3841 accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc
     3901 gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg
     3961 acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg
     4021 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     4081 gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg
     4141 tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg
     4201 tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     4261 agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg tacaagtaac
     4321 tcgagtaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4381 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4441 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4501 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4561 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4621 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4681 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4741 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4801 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4861 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcaggta
     4921 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     4981 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     5041 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5101 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5161 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5221 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5281 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5341 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5401 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     5461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5521 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5581 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     5641 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5701 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5761 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     5821 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5881 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5941 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6001 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6061 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6121 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6181 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6241 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6301 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6361 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6421 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6481 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6541 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6601 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6661 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6721 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6781 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6841 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6901 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6961 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7021 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7081 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7141 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7201 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7261 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7321 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7381 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7441 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7501 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7561 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7621 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7681 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7741 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7801 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7861 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7921 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7981 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8041 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8101 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8161 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8221 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8281 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8341 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8401 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     8461 caagctt
//
