LOCUS       V042339                 9418 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042339
VERSION     V042339
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             2317..3444
                     /label="Fmod(NM_021355)"
                     /note="Fmod(NM_021355)"
                     /gene="Fmod"
     CDS             3451..3516
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3560..3771
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3784..4052
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4081..4090
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4111..4773
                     /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             4843..4896
                     /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             4897..5496
                     /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    5497..6085
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6159..6392
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6464..6585
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6625..6760
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6793..6809)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6817..6833
                     /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(6841..6871)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6886..6907
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7195..7783)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7954..8814)
                     /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(8815..8919)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9393..9409
                     /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 ctgatacgaa ctcggaattc gccaccatgc agtgggcctc cgtcctgctg
     2341 ctggctgggc tctgctcact ttcccagggc cagtatgatg aagactctca ctggtggatc
     2401 caatacctcc gaaaccagca gtccacctac tacgacccct atgaccctta cccctatgag
     2461 ccctctgagc cttaccccta tggagtggaa gaaggcccag cctatgccta tggtgcacca
     2521 cctccaccag agccccgtga ttgtccccaa gaatgcgact gtccccccaa cttccccaca
     2581 gccatgtact gtgacaaccg caacctcaag tacctgcctt ttgtgccctc ccgcatgaag
     2641 tacgtctact tccagaacaa ccagatctct gccatccagg aaggtgtctt tgacaatgcc
     2701 actgggctcc tctgggtcgc tctacatggc aaccagatta ccagtgacaa ggtaggcagg
     2761 aaagtcttct ccaagctgag gcacttggag aggctgtact tggaccacaa caacctgacc
     2821 cggatgcccg gcccgctgcc ccgatccctg agagagctcc acctggacca caaccagatc
     2881 tcacgggtcc ccaacaatgc tttggagggc ctggagaacc tcacggcctt atatctccac
     2941 cacaatgaga tccaggaagt ggggagttcc atgagaggcc tccggtccct gatcctacta
     3001 gacctgagtt ataaccacct tcggagggta cccgacggtc tgccctcagc cctggagcag
     3061 ctgtacctag aacacaacaa tgtctacacc gtccctgaca gctacttccg ggggtcaccc
     3121 aagctgctgt acgtccggtt gtctcacaac agtctcacta acaacggcct tgctaccaac
     3181 accttcaact ccagcagcct tcttgagcta gacttgtctt acaatcagct gcagaagatc
     3241 cctcctgtca acaccaacct ggagaatctt tacctccagg gcaacaggat caatgagttc
     3301 tccatcagca gcttctgcac ggtggtggac gtcatgaact tctccaagct gcaggtgcta
     3361 cgcctggatg ggaacgagat caagcgcagc gcaatgccgg tggacgcccc actctgcctg
     3421 cgcctcgcca acctcatcga gatcggatcc gactacaaag accatgacgg tgattataaa
     3481 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat
     3541 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3601 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3661 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3721 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     3781 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     3841 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     3901 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     3961 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     4021 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac
     4081 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc
     4141 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag
     4201 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg
     4261 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag
     4321 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac
     4381 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc
     4441 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag
     4501 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg
     4561 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac
     4621 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg
     4681 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac
     4741 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct
     4801 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt
     4861 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg
     4921 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     4981 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     5041 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5101 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5161 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5221 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5281 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5341 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5401 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5461 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt
     5521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5881 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     6001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     6061 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     6121 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     6181 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     6241 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6301 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6361 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6421 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     6481 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6541 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     6601 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6661 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6721 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa
     6781 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6841 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6901 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6961 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7021 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7081 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7141 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7201 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7261 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7321 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7381 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7441 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7501 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7561 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7621 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7681 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7741 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7801 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7861 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7921 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7981 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8041 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8101 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8161 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8221 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8281 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8341 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8401 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8461 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8521 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8581 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8641 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8701 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8761 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8821 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8881 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8941 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9001 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9061 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9121 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9181 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9241 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9301 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9361 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
