LOCUS       V030705                 9520 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030705
VERSION     V030705
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3018
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3055..4374
                     /label="SQSTM1(NM_003900)"
                     /note="SQSTM1(NM_003900)"
                     /gene="SQSTM1"
     CDS             4381..4446
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4490..4701
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4714..4982
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5009..5608
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5615..6203
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6277..6510
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6582..6703
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6722..6857
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6895..6911)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6919..6935
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6943..6973)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6988..7009
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7297..7885)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8056..8916)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8917..9021)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9495..9511
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc 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 acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggcg
     3061 tcgctcaccg tgaaggccta ccttctgggc aaggaggacg cggcgcgcga gattcgccgc
     3121 ttcagcttct gctgcagccc cgagcctgag gcggaagccg aggctgcggc gggtccggga
     3181 ccctgcgagc ggctgctgag ccgggtggcc gccctgttcc ccgcgctgcg gcctggcggc
     3241 ttccaggcgc actaccgcga tgaggacggg gacttggttg ccttttccag tgacgaggaa
     3301 ttgacaatgg ccatgtccta cgtgaaggat gacatcttcc gaatctacat taaagagaaa
     3361 aaagagtgcc ggcgggacca ccgcccaccg tgtgctcagg aggcgccccg caacatggtg
     3421 caccccaatg tgatctgcga tggctgcaat gggcctgtgg taggaacccg ctacaagtgc
     3481 agcgtctgcc cagactacga cttgtgtagc gtctgcgagg gaaagggctt gcaccggggg
     3541 cacaccaagc tcgcattccc cagccccttc gggcacctgt ctgagggctt ctcgcacagc
     3601 cgctggctcc ggaaggtgaa acacggacac ttcgggtggc caggatggga aatgggtcca
     3661 ccaggaaact ggagcccacg tcctcctcgt gcaggggagg cccgccctgg ccccacggca
     3721 gaatcagctt ctggtccatc ggaggatccg agtgtgaatt tcctgaagaa cgttggggag
     3781 agtgtggcag ctgcccttag ccctctgggc attgaagttg atatcgatgt ggagcacgga
     3841 gggaaaagaa gccgcctgac ccccgtctct ccagagagtt ccagcacaga ggagaagagc
     3901 agctcacagc caagcagctg ctgctctgac cccagcaagc cgggtgggaa tgttgagggc
     3961 gccacgcagt ctctggcgga gcagatgagg aagatcgcct tggagtccga ggggcgccct
     4021 gaggaacaga tggagtcgga taactgttca ggaggagatg atgactggac ccatctgtct
     4081 tcaaaagaag tggacccgtc tacaggtgaa ctccagtccc tacagatgcc agaatccgaa
     4141 gggccaagct ctctggaccc ctcccaggag ggacccacag ggctgaagga agctgccttg
     4201 tacccacatc tcccgccaga ggctgacccg cggctgattg agtccctctc ccagatgctg
     4261 tccatgggct tctctgatga aggcggctgg ctcaccaggc tcctgcagac caagaactat
     4321 gacatcggag cggctctgga caccatccag tattcaaagc atcccccgcc gttgggatcc
     4381 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4441 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4501 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4561 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4621 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4681 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4741 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4801 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4861 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4921 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4981 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     5041 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5101 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5161 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5221 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5281 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5341 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5401 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5461 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5521 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5581 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     5641 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5701 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5761 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5821 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5881 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5941 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6001 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6061 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6121 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6181 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6241 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6301 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6361 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6421 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6481 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6541 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6721 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6781 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6841 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6901 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7081 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7981 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8041 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8101 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8161 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8221 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8281 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8341 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8401 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8461 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8521 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8581 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8641 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8701 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8761 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8821 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8881 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8941 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9001 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9061 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     9121 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9181 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9241 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9301 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9361 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9421 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9481 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
