LOCUS       V043292                 9913 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043292
VERSION     V043292
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..4767
                     /label="SUSD2(NM_019601)"
                     /note="SUSD2(NM_019601)"
                     /gene="SUSD2"
     CDS             4774..4839
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4883..5094
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5107..5375
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5402..6001
                     /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    6008..6596
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6670..6903
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6975..7096
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7115..7250
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7288..7304)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7312..7328
                     /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(7336..7366)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7381..7402
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7690..8278)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8449..9309)
                     /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(9310..9414)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9888..9904
                     /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 catgaagcca gccctcctgc cctgggccct gctgctgctg
     2341 gcgacagccc tcggcccggg ccccggaccc acagcagatg cccaagagag ctgctccatg
     2401 cgctgtggcg ccctggacgg gccatgttcc tgccacccga cgtgctctgg ccttggcacc
     2461 tgctgcttgg atttccggga cttctgcctg gagatattgc cctactcagg atccatgatg
     2521 ggcggcaagg actttgtggt gcggcacttc aagatgtcca gccccacaga cgccagtgtg
     2581 atctgcaggt ttaaggacag catccagacc ctcggccatg tggactcctc cgggcaagtg
     2641 cactgtgtgt cacctctgct ctatgagagc ggccgcatcc ccttcactgt gtcactggac
     2701 aacggccact ccttccctcg tgcgggcacc tggctggctg tgcaccccaa caaagtgtca
     2761 atgatggaga agagcgagtt ggtgaacgag acgcgttggc aatactacgg caccgccaac
     2821 acctcaggca acctcagcct gacctggcat gtcaagtcgc tgcccacgca gaccatcacc
     2881 atcgaactgt ggggctacga ggagacagga atgccctact cacaggagtg gactgcaaag
     2941 tggtcgtacc tgtaccccct ggccacacac atccccaact ccggctcttt cactttcacc
     3001 ccaaaacctg ctcctcccag ctaccagaga tggcgagtgg gtgcacttcg gatcatcgac
     3061 agcaaaaatt acgcagggca gaaggacgtg caggcgctct ggaccaacga ccacgcactg
     3121 gcctggcacc tgagcgatga cttccgagag gaccctgtgg cctgggcacg aactcagtgc
     3181 caggcctggg aggagctgga ggatcagctg cccaacttcc tggaggagct gccggactgc
     3241 ccctgcaccc tgacccaggc ccgggctgac tccggccgct tcttcacgga ctacggctgt
     3301 gacatggagc agggcagcgt gtgcacctac caccccgggg ccgtgcactg tgtgcgttct
     3361 gtgcaggcca gcctccggta cggctcaggt cagcagtgct gctacacagc ggacgggacg
     3421 cagctcctga cagctgactc cagcggcggc agcactcccg accgcggcca tgactggggc
     3481 gcacccccgt tccgcacgcc accccgagtg cccagcatgt cccactggct ctacgatgtc
     3541 ctcagcttct attactgctg cctctgggca cccgactgcc cccgctacat gcaacggcgg
     3601 ccctccaatg actgccgcaa ctaccggccc ccaagactgg cctccgcctt cggagaccca
     3661 cactttgtga ccttcgacgg caccaacttc acattcaatg ggcgcggaga gtacgtgctg
     3721 ctggaggcag cgctgaccga cctgagggtg caggcgcggg cccagcccgg gacgatgtcc
     3781 aacggcacgg agacccgtgg cactgggctg accgcagtgg ccgtccagga gggcaactca
     3841 gatgtggtgg aagtcaggct ggccaacagg accggaggtc tggaggtgct gctgaaccag
     3901 gaggtgctga gcttcaccga gcagagctgg atggacctga aaggaatgtt cctgtcggtg
     3961 gctgccgggg acagggtctc catcatgctg gcatcagggg ccggcctgga ggtcagcgtg
     4021 cagggcccgt tcctgagtgt gtccgtcctg ctgcctgaga agttcctcac ccacacccac
     4081 ggcctcctcg ggacactcaa caacgacccc accgacgact tcaccctgca cagcgggcgc
     4141 gtcctgcccc caggcaccag tccccaggag ctgttcctgt ttggggccaa ctggaccgtg
     4201 cacaatgcgt cctccctgct cacctacgat tcctggttcc tggtccacaa cttcctgtac
     4261 caacccaagc acgaccccac cttcgagccc ctcttcccca gtgagaccac cctcaacccc
     4321 agcctggcac aagaggcagc caaactatgt ggggacgatc atttctgcaa ctttgatgtg
     4381 gcagccactg ggagcctgag cacgggcact gccactcggg tggcccacca gctgcaccag
     4441 cgtcgcatgc agagcctgca gccagtggtg tcctgtggct ggctggcccc acctcccaac
     4501 ggacaaaagg agggcaacag gtacctggcg ggttccacca tctacttcca ctgtgacaac
     4561 ggctacagcc tggccggggc agagaccagc acctgccagg ctgacggcac ctggtcctca
     4621 cccaccccga agtgccagcc aggacgcagc tacgcggtgc tgttgggcat catctttggg
     4681 ggcctcgcgg tggtggcggc ggttgcgctc gtctatgtgc tgctgcgccg caggaagggc
     4741 aacacgcacg tctggggtgc acagcccgga tccgactaca aagaccatga cggtgattat
     4801 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc
     4861 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     4921 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     4981 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     5041 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     5101 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     5161 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     5221 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     5281 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     5341 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     5401 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     5461 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     5521 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     5581 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     5641 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5701 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5761 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5821 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     5881 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5941 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     6001 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     6061 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     6121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     6181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     6241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     6301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     6421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6541 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6601 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6661 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     6721 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6781 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6841 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6901 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6961 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7021 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7081 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     7141 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     7201 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     7261 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8101 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     8161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9421 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     9481 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     9541 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9601 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9661 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9721 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9781 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9841 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9901 cagtgccaag ctg
//
