LOCUS       V046925                 9516 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046925
VERSION     V046925
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3535
                     /label="Serpina1a(NM_009243)"
                     /note="Serpina1a(NM_009243)"
                     /gene="Serpina1a"
     CDS             3542..3631
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3675..3886
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3899..4167
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4196..4205
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4226..4888
                     /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             4958..5011
                     /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             5012..5611
                     /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    5612..6199
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6273..6506
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6578..6699
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6718..6853
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6891..6907)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6915..6931
                     /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(6939..6969)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6984..7005
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7293..7881)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8052..8912)
                     /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(8913..9017)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9491..9507
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatga ctccctccat ctcatggggt ctactgcttc tggcaggcct
     2341 gtgttgcctg gtccccagct ttctggctga ggatgttcag gagacagaca cctcccagaa
     2401 ggatcagtcc ccagcctccc atgagatcgc tacaaacctg ggagactttg caatcagcct
     2461 ataccgggag ctggtccatc agtccaacac ttccaacatc ttcttctccc cagtgagcat
     2521 tgccacagcc tttgctatgc tctccctagg gagcaagggt gacactcaca cgcagatcct
     2581 agagggcctg cagttcaacc tcacacaaac atcggaggct gacatccaca agtccttcca
     2641 acacctcctc caaaccctca acagaccaga cagtgagctg cagctgagca caggcaatgg
     2701 cctctttgtc aacaatgacc tgaagctggt ggagaagttt ctggaagagg ccaagaacca
     2761 ttatcaggca gaagtcttct ctgtcaactt tgcagagtca gaggaggcca agaaagtgat
     2821 taatgatttt gtggagaagg gaacccaagg aaagatagct gaggcggtga aaaaactgga
     2881 ccaagacaca gttttcgccc tggcaaatta cattctcttt aaaggcaaat ggaagaagcc
     2941 attcgatcct gagaacactg aggaagctga gttccacgtg gacgagtcca ccacggtgaa
     3001 ggtgcccatg atgaccctct cgggcatgct tcatgtgcac cattgcagca cgctgtccag
     3061 ctgggtgctg ctgatggatt atgcaggcaa tgccactgct gtcttccttc tgcccgatga
     3121 tgggaagatg cagcatctgg agcaaactct cagcaaggag ctcatctcta agttcctgct
     3181 aaacaggcgc agaaggttag cccagatcca cttccccaga ctgtccatct ctggagaata
     3241 taacttgaag acactcatga gtccactggg catcacccgg atcttcaaca atggggctga
     3301 cctctccgga atcacagagg aaaatgctcc cctgaagctc agccaggctg tgcataaggc
     3361 tgtgctgacc atcgatgaga caggaacaga agctgcagca gttacagtct tacaaatggt
     3421 tcctatgtct atgcccccta tcctgcgctt cgaccaccct ttccttttca taatatttga
     3481 agaacacact cagagcccca tctttttggg aaaagtggta gatcccacac ataaaggatc
     3541 cgaacaaaaa ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt
     3601 agagcaaaag ttaatttccg aggaggactt atagcggccg cgaaggatct gcgatcgctc
     3661 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     3721 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     3781 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     3841 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     3901 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     3961 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     4021 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     4081 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     4141 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac
     4201 catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac
     4261 cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg
     4321 catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag
     4381 ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc
     4441 cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga
     4501 cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga
     4561 cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat
     4621 ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag
     4681 cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca
     4741 catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc
     4801 cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca
     4861 cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt
     4921 ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac
     4981 atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct
     5041 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     5101 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     5161 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5221 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5281 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5341 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5401 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5461 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5521 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5581 ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga
     5641 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6001 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6121 gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc
     6181 cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     6241 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     6301 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6361 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6421 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6481 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6541 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6841 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     6901 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6961 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     9061 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     9121 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     9181 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     9241 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     9301 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     9361 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9421 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9481 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
