LOCUS       Exported                9926 bp ds-DNA     circular SYN 30-JAN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9926)
  AUTHORS   Pzx
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 26, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9926
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2224..3405
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3396
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3427..3502
                     /label=Primer 2
     regulatory      3447..3456
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3459..3479
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3480..3980
                     /label=YAPN
     misc_feature    3999..4718
                     /label=mCherry
     CDS             4719..6212
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     misc_feature    6316..6904
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6787..6798)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6979..7212
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        7417..7521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7522..8382
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8553..9141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9552..9573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9588..9618
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9626..9642
                     /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     9650..9666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac gcccctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     2581 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     2641 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     2701 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     2761 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     2821 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     2881 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     2941 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     3001 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     3061 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     3121 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     3181 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     3241 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     3301 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     3361 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaagcgg ccgcaccggt
     3421 ctgcaggcgg ccgcaccggt ctgcaggcca ccatggaccc caagaagaag aggaaggtga
     3481 tggaccccgg gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt
     3541 cgcagccccc gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga
     3601 cccaggcggc gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact
     3661 cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc
     3721 cccagaccgt gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc
     3781 ccaaatccca ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac
     3841 agcatgttcg agctcattcc tctccagctt ctctgcagtt gggagctgtt tctcctggga
     3901 cactgacccc cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc
     3961 gacagtcttc ttttgagata ctcgagtcta gaggatccat ggtgtctaaa ggcgaggagg
     4021 ataacatggc tataatcaag gaatttatga ggttcaaggt gcatatggag ggatctgtga
     4081 acggtcacga gttcgaaatc gagggcgaag gcgaggggcg accctatgaa gggacacaga
     4141 cggccaaact gaaggtgacc aagggtggcc ccctgccttt cgcctgggac atcctgtcac
     4201 cccagtttat gtacggctct aaggcttacg ttaagcaccc tgccgatatt ccggactact
     4261 tgaaactgag cttccctgaa gggttcaagt gggaacgggt aatgaatttt gaggatggcg
     4321 gtgtagtcac agttacccag gacagctctc ttcaggacgg agaatttatc tataaggtta
     4381 aacttcgggg cactaacttc ccatcagacg gccccgtgat gcagaagaaa actatgggct
     4441 gggaagccag ctcagaacgc atgtatcccg aggacggagc actgaaggga gaaatcaagc
     4501 agcgactcaa gctgaaggat gggggacatt atgatgctga ggtcaaaacc acctataagg
     4561 ccaagaaacc cgttcagctt cctggggcat acaacgtgaa tatcaaactg gatattacat
     4621 cccataacga agactatacc atcgtggaac agtacgagcg ggccgagggc agacatagca
     4681 cagggggcat ggatgaattg tacaaggcgg ccacgcgtat gaagatggac aaaaagacta
     4741 tagtttggtt tagaagagac ctaaggattg aggataatcc tgcattagca gcagctgctc
     4801 acgaaggatc tgtttttcct gtcttcattt ggtgtcctga agaagaagga cagttttatc
     4861 ctggaagagc ttcaagatgg tggatgaaac aatcacttgc tcacttatct caatccttga
     4921 aggctcttgg atctgacctc actttaatca aaacccacaa cacgatttca gcgatcttgg
     4981 attgtatccg cgttaccggt gctacaaaag tcgtctttaa ccacctctat gatcctgttt
     5041 cgttagttcg ggaccatacc gtaaaggaga agctggtgga acgtgggatc tctgtgcaaa
     5101 gctacaatgg agatctattg tatgaaccgt gggagatata ctgcgaaaag ggcaaacctt
     5161 ttacgagttt caattcttac tggaagaaat gcttagatat gtcgattgaa tccgttatgc
     5221 ttcctcctcc ttggcggttg atgccaataa ctgcagcggc tgaagcgatt tgggcgtgtt
     5281 cgattgaaga actagggctg gagaatgagg ccgagaaacc gagcaatgcg ttgttaacta
     5341 gagcttggtc tccaggatgg agcaatgctg ataagttact aaatgagttc atcgagaagc
     5401 agttgataga ttatgcaaag aacagcaaga aagttgttgg gaattctact tcactacttt
     5461 ctccgtatct ccatttcggg gaaataagcg tcagacacgt tttccagtgt gcccggatga
     5521 aacaaattat atgggcaaga gataagaaca gtgaaggaga agaaagtgca gatctttttc
     5581 ttaggggaat cggtttaaga gagtattctc ggtatatatg tttcaacttc ccgtttactc
     5641 acgagcaatc gttgttgagt catcttcggt ttttcccttg ggatgctgat gttgataagt
     5701 tcaaggcctg gagacaaggc aggaccggtt atccgttggt ggatgccgga atgagagagc
     5761 tttgggctac cggatggatg cataacagaa taagagtgat tgtttcaagc tttgctgtga
     5821 agtttcttct ccttccatgg aaatggggaa tgaagtattt ctgggataca cttttggatg
     5881 ctgatttgga atgtgacatc cttggctggc agtatatctc tgggagtatc cccgatggcc
     5941 acgagcttga tcgcttggac aatcccgcgt tacaaggcgc caaatatgac ccagaaggtg
     6001 agtacataag gcaatggctt cccgagcttg cgagattgcc aactgaatgg atccatcatc
     6061 catgggacgc tcctttaacc gtactcaaag cttctggtgt ggaactcgga acaaactatg
     6121 cgaaacccat tgtagacatc gacacagctc gtgagctact agctaaagct atttcaagaa
     6181 cccgtgaagc acagatcatg atcggagcag cagcccggga tccaccggtc gccaccggtt
     6241 cagggagcgg atccggctca gggagtgcgg ccgcaactcc cacctgcaac atgcgtgact
     6301 gaatcgatag atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6361 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6421 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6481 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6541 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6601 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6661 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     6721 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6781 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6841 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6901 ccgcctgaga tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6961 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     7021 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     7081 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7141 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7201 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7261 aatgaatatc agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga
     7321 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     7381 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     7441 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7501 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     7621 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     7681 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     7741 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     7801 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7921 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     8041 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     8101 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     8161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8221 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     8281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8341 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     8401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     8581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     8641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     8701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     8761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     9001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     9061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     9121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     9181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     9241 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     9301 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     9361 ccgattcatt aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg
     9421 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     9481 gcttttgcaa aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag
     9541 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     9601 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     9661 tatgacatga ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     9721 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     9781 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     9841 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     9901 taaatatgta ctacaaactt agtagt
//
