LOCUS       V043727                 8894 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043727
VERSION     V043727
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..3813
                     /label="SHMT2(NM_005412)"
                     /note="SHMT2(NM_005412)"
                     /gene="SHMT2"
     promoter        3864..4075
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4088..4356
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4383..4982
                     /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    4989..5577
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5651..5884
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5956..6077
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6096..6231
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6269..6285)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6293..6309
                     /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(6317..6347)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6362..6383
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6671..7259)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7430..8290)
                     /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(8291..8395)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8869..8885
                     /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 catgctgtac ttctctttgt tttgggcggc tcggcctctg
     2341 cagagatgtg ggcagctggt caggatggcc attcgggctc agcacagcaa cgcagcccag
     2401 actcagactg gggaagcaaa caggggctgg acaggccagg agagcctgtc ggacagtgat
     2461 cctgagatgt gggagttgct gcagagggag aaggacaggc agtgtcgtgg cctggagctc
     2521 attgcctcag agaacttctg cagccgagct gcgctggagg ccctggggtc ctgtctgaac
     2581 aacaagtact cggagggtta tcctggcaag agatactatg ggggagcaga ggtggtggat
     2641 gaaattgagc tgctgtgcca gcgccgggcc ttggaagcct ttgacctgga tcctgcacag
     2701 tggggagtca atgtccagcc ctactccggg tccccagcca acctggccgt ctacacagcc
     2761 cttctgcaac ctcacgaccg gatcatgggg ctggacctgc ccgatggggg ccatctcacc
     2821 cacggctaca tgtctgacgt caagcggata tcagccacgt ccatcttctt cgagtctatg
     2881 ccctataagc tcaaccccaa aactggcctc attgactaca accagctggc actgactgct
     2941 cgacttttcc ggccacggct catcatagct ggcaccagcg cctatgctcg cctcattgac
     3001 tacgcccgca tgagagaggt gtgtgatgaa gtcaaagcac acctgctggc agacatggcc
     3061 cacatcagtg gcctggtggc tgccaaggtg attccctcgc ctttcaagca cgcggacatc
     3121 gtcaccacca ctactcacaa gactcttcga ggggccaggt cagggctcat cttctaccgg
     3181 aaaggggtga aggctgtgga ccccaagact ggccgggaga tcccttacac atttgaggac
     3241 cgaatcaact ttgccgtgtt cccatccctg caggggggcc cccacaatca tgccattgct
     3301 gcagtagctg tggccctaaa gcaggcctgc acccccatgt tccgggagta ctccctgcag
     3361 gttctgaaga atgctcgggc catggcagat gccctgctag agcgaggcta ctcactggta
     3421 tcaggtggta ctgacaacca cctggtgctg gtggacctgc ggcccaaggg cctggatgga
     3481 gctcgggctg agcgggtgct agagcttgta tccatcactg ccaacaagaa cacctgtcct
     3541 ggagaccgaa gtgccatcac accgggcggc ctgcggcttg gggccccagc cttaacttct
     3601 cgacagttcc gtgaggatga cttccggaga gttgtggact ttatagatga aggggtcaac
     3661 attggcttag aggtgaagag caagactgcc aagctccagg atttcaaatc cttcctgctt
     3721 aaggactcag aaacaagtca gcgtctggcc aacctcaggc aacgggtgga gcagtttgcc
     3781 agggccttcc ccatgcctgg ttttgatgag cattgaggat ccgcggccgc gaaggatctg
     3841 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3901 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     3961 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     4021 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     4081 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     4141 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     4201 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     4261 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     4321 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     4381 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     4441 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     4501 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4561 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4621 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4681 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4741 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4801 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4861 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4921 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4981 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     5041 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5101 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5161 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5221 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     5281 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     5341 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     5401 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     5461 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     5521 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     5581 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5641 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt
     5701 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5761 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5821 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5881 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     5941 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6001 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6061 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt
     6121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6181 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt
     6241 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6301 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6361 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6421 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6481 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6541 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6601 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6661 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6721 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6781 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6841 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6901 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6961 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7021 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7081 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7141 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7201 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7261 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7321 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7381 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7441 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7501 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7561 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7621 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7681 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7741 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7801 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7861 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7921 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7981 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8041 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8101 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8161 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8221 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8281 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8341 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8401 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     8461 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     8521 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     8581 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     8641 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     8701 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     8761 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     8821 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     8881 ccagtgccaa gctg
//
