LOCUS       V024334                 8935 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024334
VERSION     V024334
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..3828
                     /label="SLC2A4(NM_001042)"
                     /note="SLC2A4(NM_001042)"
                     /gene="SLC2A4"
     CDS             3835..3861
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3905..4116
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4129..4397
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4424..5023
                     /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    5030..5618
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5692..5925
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5997..6118
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6137..6272
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6310..6326)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6334..6350
                     /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(6358..6388)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6403..6424
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6712..7300)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7471..8331)
                     /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(8332..8436)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8910..8926
                     /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 catgccgtcg ggcttccaac agataggctc cgaagatggg
     2341 gaaccccctc agcagcgagt gactgggacc ctggtccttg ctgtgttctc tgcggtgctt
     2401 ggctccctgc agtttgggta caacattggg gtcatcaatg cccctcagaa ggtgattgaa
     2461 cagagctaca atgagacgtg gctggggagg caggggcctg agggacccag ctccatccct
     2521 ccaggcaccc tcaccaccct ctgggccctc tccgtggcca tcttttccgt gggcggcatg
     2581 atttcctcct tcctcattgg tatcatctct cagtggcttg gaaggaaaag ggccatgctg
     2641 gtcaacaatg tcctggcggt gctggggggc agcctcatgg gcctggccaa tgctgctgcc
     2701 tcctatgaaa tgctcatcct tggacgattc ctcattggcg cctactcagg gctgacatca
     2761 gggctggtgc ccatgtacgt gggggagatt gctcccactc acctgcgggg cgccctgggg
     2821 acgctcaacc aactggccat tgttatcggc attctgatcg cccaggtgct gggcttggag
     2881 tccctcctgg gcactgccag cctgtggcca ctgctcctgg gcctcacagt gctacctgcc
     2941 ctcctgcagc tggtcctgct gcccttctgt cccgagagcc cccgctacct ctacatcatc
     3001 cagaatctcg aggggcctgc cagaaagagt ctgaagcgcc tgacaggctg ggccgatgtt
     3061 tctggagtgc tggctgagct gaaggatgag aagcggaagc tggagcgtga gcggccactg
     3121 tccctgctcc agctcctggg cagccgtacc caccggcagc ccctgatcat tgcggtcgtg
     3181 ctgcagctga gccagcagct ctctggcatc aatgctgttt tctattattc gaccagcatc
     3241 ttcgagacag caggggtagg ccagcctgcc tatgccacca taggagctgg tgtggtcaac
     3301 acagtcttca ccttggtctc ggtgttgttg gtggagcggg cggggcgccg gacgctccat
     3361 ctcctgggcc tggcgggcat gtgtggctgt gccatcctga tgactgtggc tctgctcctg
     3421 ctggagcgag ttccagccat gagctacgtc tccattgtgg ccatctttgg cttcgtggca
     3481 ttttttgaga ttggccctgg ccccattcct tggttcatcg tggccgagct cttcagccag
     3541 ggaccccgcc cggcagccat ggctgtggct ggtttctcca actggacgag caacttcatc
     3601 attggcatgg gtttccagta tgttgcggag gctatggggc cctacgtctt ccttctattt
     3661 gcggtcctcc tgctgggctt cttcatcttc accttcttaa gagtacctga aactcgaggc
     3721 cggacgtttg accagatctc agctgccttc caccggacac cctctctttt agagcaggag
     3781 gtgaaaccca gcacagaact tgagtattta gggccagatg agaacgacgg atcctaccca
     3841 tacgacgtac cagattacgc ttagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3901 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3961 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     4021 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     4081 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     4141 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     4201 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     4261 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     4321 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     4381 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     4441 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     4501 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     4561 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4621 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4681 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4741 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4801 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4861 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4921 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4981 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     5041 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5101 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5161 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5221 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5281 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5341 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5401 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5461 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5521 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5581 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     5641 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5701 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     5761 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5821 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5881 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     5941 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     6001 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6061 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6121 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6181 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6241 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     6301 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6361 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6421 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6481 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7141 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8461 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8521 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     8581 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8641 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     8701 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     8761 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     8821 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     8881 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
