LOCUS       V043233                 9325 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043233
VERSION     V043233
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..4179
                     /label="NXF2(NM_022053)"
                     /note="NXF2(NM_022053)"
                     /gene="NXF2"
     CDS             4186..4251
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4295..4506
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4519..4787
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4814..5413
                     /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    5420..6008
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6082..6315
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6387..6508
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6527..6662
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6700..6716)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6724..6740
                     /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(6748..6778)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6793..6814
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7102..7690)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7861..8721)
                     /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(8722..8826)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9300..9316
                     /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 catgtgctct actctaaaga agtgtgggac atacagaact
     2341 gaagttgcag aatgccatga ccatggtagc acttttcaag gaagaaagaa aggtgggagt
     2401 tctttccggg ataattttga caagaggagc tgtcattatg aacatggtgg gtatgagcgc
     2461 ccgccttcac actgccagga gaatgatgga agcgtggaga tgagggatgt ccacaaggac
     2521 caacaactaa gacacactcc ttatagcatc cgatgcgaaa gaagaatgaa atggcatagt
     2581 gaagacgaaa tccgtattac cacgtggaga aatagaaaac ctccggagag aaaaatgagt
     2641 cagaacacac aggatggata cacaaggaac tggtttaagg tcacaattcc ttacgggata
     2701 aagtatgaca aggcatggct aatgaattca atccagagcc attgcagtga ccgcttcact
     2761 ccggttgatt tccactacgt ccgaaatcgg gcatgcttct ttgtccagga tgctagcgct
     2821 gcctccgcat tgaaggatgt cagttataag atttatgatg atgagaacca aaagatatgt
     2881 atatttgtca atcattctac tgcgccctac tctgtgaaga ataagttgaa gccaggccaa
     2941 atggagatgc taaagctgac catgaacaaa cggtacaatg tctcccagca agctcttgat
     3001 ctccagaatc tccgctttga cccagacttg atgggccgtg acattgatat aatcctgaat
     3061 cgaagaaact gcatggctgc caccctgaag atcattgaaa gaaatttccc tgagctgttg
     3121 tctttgaact tgtgcaacaa caagctgtac cagctggatg gcctttctga cattacagag
     3181 aaggctccca aagtcaagac cctgaatctc tccaaaaata agctggagtc ggcgtgggag
     3241 ttgggcaagg tgaaagggct gaagctcgaa gagctatggc tagaagggaa cccgttgtgc
     3301 agcaccttct cggaccagtc cgcctatgta agtgccatcc gggattgttt ccccaagttg
     3361 ttacgcctgg acggccgaga gttatccgca ccagtgattg ttgacattga cagctctgag
     3421 acaatgaaac cctgcaagga aaactttact ggatctgaga ccctaaagca tttagtcctg
     3481 caattcctgc agcagtatta ctcgatctat gactctggag atcgacaggg tctcctcggt
     3541 gcttaccacg atgaggcctg cttctccttg gctattccct tcgaccccaa ggactcagcc
     3601 ccgagcagct tgtgcaagta ctttgaggat agcaggaata tgaaaacact caaggacccc
     3661 tacctgaagg gggaactgct gaggcgcaca aaacgtgaca ttgtggactc cctcagtgcg
     3721 ttgcccaaaa ctcagcatga cctcagctcc atcctggtgg acgtgtggtg ccagacggaa
     3781 aggatgctct gcttttctgt caatggggtt ttcaaggaag tggaaggaca gtctcagggt
     3841 tctgttctcg ccttcacccg gaccttcatt gctacccctg gcagcagttc cagtctgtgc
     3901 atcgtgaatg acgagctgtt tgtgagggat gccagccccc aagagactca gagtgccttc
     3961 tccatcccag tgtccacact ctcctccagc tctgagccct ccctctccca ggagcagcag
     4021 gaaatggtgc aggctttctc tgcccagtct gggatgaaac tggagtggtc tcagaagtgc
     4081 cttcaggaca atgagtggaa ctacactaga gctggccagg ccttcactat gctccagacc
     4141 gagggcaaga tccccgcgga ggccttcaag caaatctccg gatccgacta caaagaccat
     4201 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg
     4261 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     4321 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     4381 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4441 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4501 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     4561 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     4621 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4681 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4741 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4801 gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     4861 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     4921 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     4981 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5041 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5101 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5161 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     5221 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5281 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5341 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     5401 gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5461 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5521 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5581 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5641 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5701 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5761 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5821 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5881 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5941 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6001 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     6061 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc
     6121 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6181 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6241 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6301 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     6361 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     6421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6481 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     6541 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6601 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6661 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg
     6721 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6781 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6841 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6901 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6961 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7021 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7081 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7141 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7201 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7261 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7321 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7381 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7441 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7501 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7561 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7621 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7681 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7741 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7801 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7861 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7921 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7981 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8041 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8101 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8161 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8221 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8281 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8341 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8401 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8461 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8521 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8581 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8641 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8701 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8761 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8821 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     8881 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     8941 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     9001 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     9061 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     9121 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     9181 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     9241 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     9301 taaaacgacg gccagtgcca agctg
//
