LOCUS       V033206                 9562 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033206
VERSION     V033206
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2843..4318
                     /label="SLC2A1(NM_006516)"
                     /note="SLC2A1(NM_006516)"
                     /gene="SLC2A1"
     promoter        4350..4849
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4870..5469
                     /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    5483..6071
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6278..6911
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7040..7056)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7064..7080
                     /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(7088..7118)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7133..7154
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7442..8030)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8201..9061)
                     /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(9062..9166)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9214..9348
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattcgcca ccatggagcc cagcagcaag aagctgacgg gtcgcctcat
     2881 gctggccgtg ggaggagcag tgcttggctc cctgcagttt ggctacaaca ctggagtcat
     2941 caatgccccc cagaaggtga tcgaggagtt ctacaaccag acatgggtcc accgctatgg
     3001 ggagagcatc ctgcccacca cgctcaccac gctctggtcc ctctcagtgg ccatcttttc
     3061 tgttgggggc atgattggct ccttctctgt gggccttttc gttaaccgct ttggccggcg
     3121 gaattcaatg ctgatgatga acctgctggc cttcgtgtcc gccgtgctca tgggcttctc
     3181 gaaactgggc aagtcctttg agatgctgat cctgggccgc ttcatcatcg gtgtgtactg
     3241 cggcctgacc acaggcttcg tgcccatgta tgtgggtgaa gtgtcaccca cagcccttcg
     3301 tggggccctg ggcaccctgc accagctggg catcgtcgtc ggcatcctca tcgcccaggt
     3361 gttcggcctg gactccatca tgggcaacaa ggacctgtgg cccctgctgc tgagcatcat
     3421 cttcatcccg gccctgctgc agtgcatcgt gctgcccttc tgccccgaga gtccccgctt
     3481 cctgctcatc aaccgcaacg aggagaaccg ggccaagagt gtgctaaaga agctgcgcgg
     3541 gacagctgac gtgacccatg acctgcagga gatgaaggaa gagagtcggc agatgatgcg
     3601 ggagaagaag gtcaccatcc tggagctgtt ccgctccccc gcctaccgcc agcccatcct
     3661 catcgctgtg gtgctgcagc tgtcccagca gctgtctggc atcaacgctg tcttctatta
     3721 ctccacgagc atcttcgaga aggcgggggt gcagcagcct gtgtatgcca ccattggctc
     3781 cggtatcgtc aacacggcct tcactgtcgt gtcgctgttt gtggtggagc gagcaggccg
     3841 gcggaccctg cacctcatag gcctcgctgg catggcgggt tgtgccatac tcatgaccat
     3901 cgcgctagca ctgctggagc agctaccctg gatgtcctat ctgagcatcg tggccatctt
     3961 tggctttgtg gccttctttg aagtgggtcc tggccccatc ccatggttca tcgtggctga
     4021 actcttcagc cagggtccac gtccagctgc cattgccgtt gcaggcttct ccaactggac
     4081 ctcaaatttc attgtgggca tgtgcttcca gtatgtggag caactgtgtg gtccctacgt
     4141 cttcatcatc ttcactgtgc tcctggttct gttcttcatc ttcacctact tcaaagttcc
     4201 tgagactaaa ggccggacct tcgatgagat cgcttccggc ttccggcagg ggggagccag
     4261 ccaaagtgac aagacacccg aggagctgtt ccatcccctg ggggctgatt cccaagtgtg
     4321 aggatcccgc gactctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     4381 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     4441 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     4501 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     4561 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     4621 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     4681 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     4741 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     4801 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
     4861 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4921 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4981 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5041 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5101 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5161 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5221 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5281 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5341 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5401 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5461 ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     5521 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5581 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5641 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5701 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5761 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5821 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5881 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5941 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     6001 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6061 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     6121 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     6181 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     6241 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa
     6301 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag
     6361 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag
     6421 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg
     6481 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg
     6541 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag
     6601 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt
     6661 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt
     6721 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6781 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6841 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6901 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     6961 atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct
     7021 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9181 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt
     9241 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9301 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     9361 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     9421 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     9481 tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa
     9541 tatgtactac aaacttagta gt
//
