LOCUS       V039093                 9442 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039093
VERSION     V039093
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             2303..3556
                     /label="SERPINA1(NM_000295)"
                     /note="SERPINA1(NM_000295)"
                     /gene="SERPINA1"
     promoter        3601..3812
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3825..4093
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4122..4131
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4152..4814
                     /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             4884..4937
                     /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             4938..5537
                     /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    5538..6125
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6199..6432
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6504..6625
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6644..6779
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6817..6833)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6841..6857
                     /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(6865..6895)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6910..6931
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7219..7807)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7978..8838)
                     /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(8839..8943)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9417..9433
                     /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 gaattcgcca ccatgccgtc ttctgtctcg tggggcatcc tcctgctggc
     2341 aggcctgtgc tgcctggtcc ctgtctccct ggctgaggat ccccagggag atgctgccca
     2401 gaagacagat acatcccacc atgatcagga tcacccaacc ttcaacaaga tcacccccaa
     2461 cctggctgag ttcgccttca gcctataccg ccagctggca caccagtcca acagcaccaa
     2521 tatcttcttc tccccagtga gcatcgctac agcctttgca atgctctccc tggggaccaa
     2581 ggctgacact cacgatgaaa tcctggaggg cctgaatttc aacctcacgg agattccgga
     2641 ggctcagatc catgaaggct tccaggaact cctccgtacc ctcaaccagc cagacagcca
     2701 gctccagctg accaccggca atggcctgtt cctcagcgag ggcctgaagc tagtggataa
     2761 gtttttggag gatgttaaaa agttgtacca ctcagaagcc ttcactgtca acttcgggga
     2821 caccgaagag gccaagaaac agatcaacga ttacgtggag aagggtactc aagggaaaat
     2881 tgtggatttg gtcaaggagc ttgacagaga cacagttttt gctctggtga attacatctt
     2941 ctttaaaggc aaatgggaga gaccctttga agtcaaggac accgaggaag aggacttcca
     3001 cgtggaccag gtgaccaccg tgaaggtgcc tatgatgaag cgtttaggca tgtttaacat
     3061 ccagcactgt aagaagctgt ccagctgggt gctgctgatg aaatacctgg gcaatgccac
     3121 cgccatcttc ttcctgcctg atgaggggaa actacagcac ctggaaaatg aactcaccca
     3181 cgatatcatc accaagttcc tggaaaatga agacagaagg tctgccagct tacatttacc
     3241 caaactgtcc attactggaa cctatgatct gaagagcgtc ctgggtcaac tgggcatcac
     3301 taaggtcttc agcaatgggg ctgacctctc cggggtcaca gaggaggcac ccctgaagct
     3361 ctccaaggcc gtgcataagg ctgtgctgac catcgacgag aaagggactg aagctgctgg
     3421 ggccatgttt ttagaggcca tacccatgtc tatccccccc gaggtcaagt tcaacaaacc
     3481 ctttgtcttc ttaatgattg aacaaaatac caagtctccc ctcttcatgg gaaaagtggt
     3541 gaatcccacc caaaaataag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt
     3601 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     3661 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     3721 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     3781 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     3841 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     3901 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     3961 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     4021 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     4081 gccgttacag atccaagctg tgaccggcgc ctacgctaga cgccaccatg gagagcgacg
     4141 agagcggcct gcccgccatg gagatcgagt gccgcatcac cggcaccctg aacggcgtgg
     4201 agttcgagct ggtgggcggc ggagagggca cccccaagca gggccgcatg accaacaaga
     4261 tgaagagcac caaaggcgcc ctgaccttca gcccctacct gctgagccac gtgatgggct
     4321 acggcttcta ccacttcggc acctacccca gcggctacga gaaccccttc ctgcacgcca
     4381 tcaacaacgg cggctacacc aacacccgca tcgagaagta cgaggacggc ggcgtgctgc
     4441 acgtgagctt cagctaccgc tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg
     4501 gcaccggctt ccccgaggac agcgtgatct tcaccgacaa gatcatccgc agcaacgcca
     4561 ccgtggagca cctgcacccc atgggcgata acgtgctggt gggcagcttc gcccgcacct
     4621 tcagcctgcg cgacggcggc tactacagct tcgtggtgga cagccacatg cacttcaaga
     4681 gcgccatcca ccccagcatc ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg
     4741 aggagctgca cagcaacacc gagctgggca tcgtggagta ccagcacgcc ttcaagaccc
     4801 ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg
     4861 gacccggctc caccggatct cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg
     4921 aggagaatcc cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     4981 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     5041 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     5101 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     5161 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     5221 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     5281 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     5341 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     5401 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5461 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5521 gcaagcccgg tgcctgaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5581 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5641 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5701 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5761 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5821 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5881 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     5941 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6001 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6061 ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg gatctccctt tggccgcctc
     6121 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6181 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     6241 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6301 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6361 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6421 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6481 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6541 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6601 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6661 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6721 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6781 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     6841 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6901 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6961 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7021 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7621 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8941 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     9001 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     9061 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     9121 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     9181 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     9241 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     9301 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     9361 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     9421 aacgacggcc agtgccaagc tg
//
