LOCUS       V041584                10198 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041584
VERSION     V041584
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..5052
                     /label="Hk2(NM_013820)"
                     /note="Hk2(NM_013820)"
                     /gene="Hk2"
     CDS             5059..5124
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5168..5379
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5392..5660
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5687..6286
                     /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    6293..6881
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6955..7188
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7260..7381
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7400..7535
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7573..7589)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7597..7613
                     /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(7621..7651)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7666..7687
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7975..8563)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8734..9594)
                     /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(9595..9699)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10173..10189
                     /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 catgatcgcc tcgcatatga tcgcctgctt attcacggag
     2341 ctcaaccaaa accaagtgca gaaggttgac cagtatctct accacatgcg tctctcagat
     2401 gagacccttc tggagatttc taggcggttc cggaaggaga tggagaaggg gctaggagct
     2461 accacacacc ctacagcagc tgtgaagatg ctgcccacct ttgtgaggtc aactccggat
     2521 gggacagaac atggcgagtt cctggctctg gaccttggag gaaccaactt ccgtgtgctc
     2581 cgagtaaggg tgacagacaa tggtctccag agggtggaga tggagaacca gatctacgcc
     2641 attcccgagg acatcatgcg gggcagtgga acccagctgt ttgaccacat tgccgaatgc
     2701 ctggccaact tcatggacaa gctacagatc aaagagaaga agctcccgct gggtttcacc
     2761 ttctcgttcc cctgccacca gacgaaactg gatgagagtt tcttggtctc atggactaag
     2821 gggttcaagt ccagtggcgt ggaaggcaga gatgtggtgg atctgatccg gaaggccatc
     2881 cagcgcagag gggactttga cattgacatt gtggctgtgg tgaatgacac ggttgggacc
     2941 atgatgactt gtggttatga tgatcagaac tgcgagattg gtctcattgt gggtactggc
     3001 agtaatgcct gctacatgga ggagatgcgt cacattgaca tggtggaggg cgacgaggga
     3061 cggatgtgca tcaacatgga gtggggggcc tttggggacg acggtacact caatgacatc
     3121 cgaactgagt ttgacagaga gatcgacatg ggctcactga accctgggaa gcagctgttt
     3181 gagaagatga tcagcgggat gtacatgggg gagctggtca gactcatcct ggtgaagatg
     3241 gccaaggccg agctgctgtt ccaagggaaa ctcagcccag agctccttac cactggctcc
     3301 ttcgagacca aagatgtctc ggatattgaa gacgataagg acggaattca gaaggcctac
     3361 cagatcctgg tgcgcctggg tctgagcccg ctgcaggagg actgtgtggc cacgcaccga
     3421 atctgccaga ttgtgtccac gcgctcagcc agcctgtgtg cagccaccct ggccgcggtg
     3481 ctgtggcgga tcaaagagaa caagggcgag gagcgactcc gctccaccat cggcgtggat
     3541 ggctctgtct acaagaaaca tccccatttt gccaagcgtc tccataaggc agtgaggagg
     3601 ctggtgcccg attgtgatgt ccgcttcctc cgctctgagg atggcagcgg caagggggcg
     3661 gctatggtga cggcggtggc ttaccgtctg gctgaccaac accgggcccg ccagaagacc
     3721 ctggagtctc tgaagctgag ccatgaacag ctgctggagg ttaagagaag gatgaaggtg
     3781 gaaatggagc agggtctgag caaggagacg catgaggccg cccctgtgaa gatgttgccc
     3841 acttatgtgt gtgccactcc agacggtaca gagaaaggag acttcctggc cttggatctt
     3901 ggaggaacaa acttccgggt cctgctggtg cgcgtgcgta atggcaagcg aaggggcgta
     3961 gagatgcata acaagatcta ctccatccca caggaagtca tgcatggcac tggagaagag
     4021 cttttcgacc acattgtcca gtgcatcgcg gacttccttg agtacatggg catgaagggc
     4081 gtgtccctac ctttgggttt caccttctcc ttcccttgcc agcagaacag cctggaccag
     4141 agcatcctcc tcaagtggac aaagggattc aaggcatccg gctgcgaggg tgaagatgtg
     4201 gtcaccttgc tgaaggaagc cattcgccgg cgagaggagt ttgacctgga tgtggtggcc
     4261 gtggtaaatg acacagttgg gactatgatg acttgtggct atgaagaccc tcactgtgaa
     4321 gttggcctca ttgttggcac tggaagcaat gcctgctaca tggaggagat gcgtaatgtg
     4381 gaactggtgg acggagagga ggggcggatg tgtgtcaaca tggagtgggg agcgttcggg
     4441 gacaacggct gcctggatga cctgcgcaca gtgtttgacg tcgctgtgga cgagctctct
     4501 ctcaaccctg gcaaacagag gtttgagaag atgatcagcg gcatgtactt gggcgagatt
     4561 gtgcgcaaca tcctgatcga tttcacaaag cgggggctgc tcttccgtgg ccgcatctca
     4621 gagcgcctca agacaagggg aatcttcgaa actaagttcc tgtctcagat cgagagtgac
     4681 tgcctggccc tgctacaggt ccgagccatc ttgcgccacc tagggctgga gagcacgtgt
     4741 gacgacagca tcattgtgaa ggaggtgtgc actgtggtcg cccggcgcgc ggcccagctc
     4801 tgtggcgcag gcatggccgc cgtggtggac aagataagag agaaccgtgg actggacaac
     4861 ctcaaagtga cggtgggcgt ggacgggaca ctgtacaagc ttcatcctca ctttgccaaa
     4921 gtcatgcatg agacggtgag agatctggct ccgaaatgtg acgtgtcctt cctggaatcc
     4981 gaggacggca gtgggaaggg agcggctctc atcactgctg tggcctgtcg catccgggag
     5041 gccgggcaga gaggatccga ctacaaagac catgacggtg attataaaga tcatgacatc
     5101 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc
     5161 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     5221 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     5281 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     5341 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     5401 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     5461 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     5521 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     5581 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     5641 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa
     5701 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     5761 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     5821 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5881 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5941 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     6001 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     6061 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     6121 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     6181 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     6241 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc
     6301 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     6361 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     6421 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6481 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6541 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6601 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6661 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     6721 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6781 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6841 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     6901 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6961 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     7021 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     7081 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     7141 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     7201 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     7261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7381 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     7441 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7501 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     7561 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7621 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7681 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7741 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7801 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7861 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7921 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7981 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8041 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8101 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8161 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8221 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8281 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8341 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8401 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8461 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8521 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8581 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8641 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8701 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8761 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8821 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8881 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8941 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9001 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9061 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9121 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9181 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9241 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9301 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9361 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9421 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9481 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9541 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9601 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9661 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9721 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9781 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9841 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9901 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9961 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
    10021 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
    10081 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
    10141 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
