LOCUS       V020579                 9286 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020579
VERSION     V020579
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2295..3371
                     /label="SCD(NM_005063)"
                     /note="SCD(NM_005063)"
                     /gene="SCD"
     CDS             3372..3395
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        3445..3656
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3669..3937
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3966..3975
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3996..4658
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4728..4781
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4782..5381
                     /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    5382..5969
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6043..6276
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6348..6469
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6488..6623
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6661..6677)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6685..6701
                     /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(6709..6739)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6754..6775
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7063..7651)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7822..8682)
                     /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(8683..8787)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9261..9277
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc caccatgccg gcccacttgc tgcaggacga tatctctagc tcctatacca
     2341 ccaccaccac cattacagcg cctccctcca gggtcctgca gaatggagga gataagttgg
     2401 agacgatgcc cctctacttg gaagacgaca ttcgccctga tataaaagat gatatatatg
     2461 accccaccta caaggataag gaaggcccaa gccccaaggt tgaatatgtc tggagaaaca
     2521 tcatccttat gtctctgcta cacttgggag ccctgtatgg gatcactttg attcctacct
     2581 gcaagttcta cacctggctt tggggggtat tctactattt tgtcagtgcc ctgggcataa
     2641 cagcaggagc tcatcgtctg tggagccacc gctcttacaa agctcggctg cccctacggc
     2701 tctttctgat cattgccaac acaatggcat tccagaatga tgtctatgaa tgggctcgtg
     2761 accaccgtgc ccaccacaag ttttcagaaa cacatgctga tcctcataat tcccgacgtg
     2821 gctttttctt ctctcacgtg ggttggctgc ttgtgcgcaa acacccagct gtcaaagaga
     2881 aggggagtac gctagacttg tctgacctag aagctgagaa actggtgatg ttccagagga
     2941 ggtactacaa acctggcttg ctgatgatgt gcttcatcct gcccacgctt gtgccctggt
     3001 atttctgggg tgaaactttt caaaacagtg tgttcgttgc cactttcttg cgatatgctg
     3061 tggtgcttaa tgccacctgg ctggtgaaca gtgctgccca cctcttcgga tatcgtcctt
     3121 atgacaagaa cattagcccc cgggagaata tcctggtttc acttggagct gtgggtgagg
     3181 gcttccacaa ctaccaccac tcctttccct atgactactc tgccagtgag taccgctggc
     3241 acatcaactt caccacattc ttcattgatt gcatggccgc cctcggtctg gcctatgacc
     3301 ggaagaaagt ctccaaggcc gccatcttgg ccaggattaa aagaaccgga gatggaaact
     3361 acaagagtgg cgattacaag gatgacgacg ataagtagga tccgcggccg cgaaggatct
     3421 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     3481 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     3541 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     3601 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     3661 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     3721 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     3781 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     3841 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     3901 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc
     3961 tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca
     4021 tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca
     4081 agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct
     4141 acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct
     4201 acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga
     4261 agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg
     4321 tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg
     4381 acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc
     4441 tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg
     4501 tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc
     4561 ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg
     4621 agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca
     4681 attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa
     4741 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca
     4801 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4861 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4921 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4981 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     5041 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5101 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     5161 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     5221 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     5281 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5341 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca
     5401 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5461 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5521 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5581 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5641 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5701 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5761 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     5821 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5881 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga
     5941 gtcggatctc cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     6001 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6061 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6121 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6181 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6241 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6301 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6361 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6421 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6481 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6541 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6601 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7501 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     8821 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     8881 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     8941 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     9001 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     9061 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     9121 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     9181 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     9241 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
