LOCUS       Exported                9352 bp ds-DNA     circular SYN 21-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Nov 22, 2020 from SnapGene Viewer 5.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9352
                     /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"
     promoter        complement(2369..3085)
                     /label=EGFP
     misc_feature    complement(2408..2427)
                     /label=seq primer 2
     primer_bind     2572..2590
                     /label=Primer 3-SET2
     primer_bind     complement(2572..2590)
                     /label=Primer 2
     primer_bind     complement(2593..2616)
                     /label=probe-SET2
     primer_bind     2613..2633
                     /label=qpcr rev
     primer_bind     complement(2649..2669)
                     /label=Primer 1-SET2
     primer_bind     complement(2706..2723)
                     /label=qpcr fwd
     promoter        complement(3166..3443)
                     /label=chicken beta-actin promoter
     enhancer        3445..3824
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3986..4481
                     /label=PGK
     CDS             4500..5201
                     /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    5212..5800
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5683..5694)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5873..6106
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6178..6299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6339..6474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6495..6513)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6523..6539)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6681..7136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7162..7266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7267..8127
                     /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      8298..8886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9174..9195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9210..9240
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9248..9264
                     /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     9272..9288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9309..9327
                     /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 cggccgctac cggggcgggc ggcgcgaggc
     3121 ggcggcggag cggggcacgg ggcgaaggca gcgcgcagcg actcccgccc gccgcgcgct
     3181 tcgcttttta tagggccgcc gccgccgccg cctcgccata aaaggaaact ttcggagcgc
     3241 gccgctctga ttggctgccg ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg
     3301 ccccgccccg cctggcgcgc gccccccccc cccccccgcc cccatcgctg cacaaaataa
     3361 ttaaaaaata aataaataca aaattggggg tggggagggg ggggagatgg ggagagtgaa
     3421 gcagaacgtg gggctcacct cgaccatggt aatagcgatg actaatacgt agatgtactg
     3481 ccaagtagga aagtcccgta aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     3541 gtcattgacg tcaatagggg gcggacttgg catatgatac acttgatgta ctgccaagtg
     3601 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     3661 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     3721 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     3781 ccccgtaatt gattactatt aataactagt caataatcaa tgccaacatg gcggtcatat
     3841 tggacatgag ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca
     3901 atagccaata ttgatttatg ctatataacc aatgaataat atggctaatg gccaatattg
     3961 aagatccccc gggctgcagg aattccagta cttttcccaa ggcagtctgg agcatgcgct
     4021 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     4081 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     4141 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     4201 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     4261 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     4321 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     4381 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     4441 gcgctgttct cctcttcctc atctccgggc ctttcgacct gaagcttgcc gccggcacca
     4501 tggtgagcaa gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg
     4561 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     4621 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     4681 tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc
     4741 ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     4801 aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct
     4861 acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga
     4921 ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     4981 agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca
     5041 tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg
     5101 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc
     5161 gccaccacct gttcctgtac ggcatggacg agctgtacaa gtaactcgag taatcaacct
     5221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5581 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     5641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5761 cctcagacga gtcggatctc cctttgggcc gcctccccgc aggtaccttt aagaccaatg
     5821 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     5881 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     5941 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6001 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6061 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     6121 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     6181 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6241 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6301 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6421 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga
     6481 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     6541 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     6601 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6661 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6721 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6781 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6841 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6901 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6961 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     7021 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     7081 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     7141 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7201 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7261 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7321 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7381 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7441 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7501 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7561 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7621 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7681 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7741 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7801 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7861 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7921 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7981 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8041 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8101 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8161 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8221 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8281 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8341 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8401 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8461 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8521 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8581 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8641 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8701 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8761 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8821 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8881 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8941 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     9001 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9061 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     9121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     9181 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     9241 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     9301 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc tt
//
