LOCUS       V036895                 9102 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036895
VERSION     V036895
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3289
                     /label="CTSS(NM_004079)"
                     /note="CTSS(NM_004079)"
                     /gene="CTSS"
     CDS             3311..4030
                     /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"
     promoter        4072..4283
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4296..4564
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4591..5190
                     /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    5197..5785
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5859..6092
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6164..6285
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6304..6439
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6477..6493)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6501..6517
                     /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(6525..6555)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6570..6591
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6879..7467)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7638..8498)
                     /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(8499..8603)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9077..9093
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatga aacggctggt ttgtgtgctc ttggtgtgct cctctgcagt
     2341 ggcacagttg cataaagatc ctaccctgga tcaccactgg catctctgga agaaaaccta
     2401 tggcaaacaa tacaaggaaa agaatgaaga agcagtacga cgtctcatct gggaaaagaa
     2461 tctaaagttt gtgatgcttc acaacctgga gcattcaatg ggaatgcact catacgatct
     2521 gggcatgaac cacctgggag acatgaccag tgaagaagtg atgtctttga tgagttccct
     2581 gagagttccc agccagtggc agagaaatat cacatataag tcaaacccta atcggatatt
     2641 gcctgattct gtggactgga gagagaaagg gtgtgttact gaagtgaaat atcaaggttc
     2701 ttgtggtgct tgctgggctt tcagtgctgt gggggccctg gaagcacagc tgaagctgaa
     2761 aacaggaaag ctggtgtctc tcagtgccca gaacctggtg gattgctcaa ctgaaaaata
     2821 tggaaacaaa ggctgcaatg gtggcttcat gacaacggct ttccagtaca tcattgataa
     2881 caagggcatc gactcagacg cttcctatcc ctacaaagcc atggatcaga aatgtcaata
     2941 tgactcaaaa tatcgtgctg ccacatgttc aaagtacact gaacttcctt atggcagaga
     3001 agatgtcctg aaagaagctg tggccaataa aggcccagtg tctgttggtg tagatgcgcg
     3061 tcatccttct ttcttcctct acagaagtgg tgtctactat gaaccatcct gtactcagaa
     3121 tgtgaatcat ggtgtacttg tggttggcta tggtgatctt aatgggaaag aatactggct
     3181 tgtgaaaaac agctggggcc acaactttgg tgaagaagga tatattcgga tggcaagaaa
     3241 taaaggaaat cattgtggga ttgctagctt tccctcttac ccagaaatcg gatccggagg
     3301 agggggaagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3361 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3421 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3481 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3541 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3601 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3661 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3721 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3781 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3841 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3901 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3961 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4021 gtacaagtaa gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc
     4081 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct
     4141 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     4201 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     4261 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg
     4321 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg
     4381 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac
     4441 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt
     4501 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca
     4561 gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc
     4621 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4681 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     4741 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4801 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4861 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4921 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4981 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5041 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5101 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5161 tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt
     5221 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5281 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5341 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5401 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5461 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5521 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5581 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5641 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5701 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5761 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     5821 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     5881 acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     5941 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6001 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6061 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6121 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     6181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     6301 gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6361 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6421 ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca
     6481 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6541 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6601 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6661 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6721 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6781 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6841 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6901 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6961 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7021 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7081 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7141 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7201 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7261 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7321 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7381 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7441 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7501 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7561 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7621 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7681 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7741 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7801 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7861 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7921 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7981 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8041 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8101 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8161 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8221 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8281 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8341 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8401 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8461 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8521 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8581 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8641 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     8701 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     8761 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     8821 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     8881 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     8941 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     9001 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     9061 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
