LOCUS       V032130                 9508 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032130
VERSION     V032130
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..3550
                     /label="Serpina3k(NM_011458)"
                     /note="Serpina3k(NM_011458)"
                     /gene="Serpina3k"
     CDS             3557..3622
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3667..3878
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3891..4159
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4188..4197
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4218..4880
                     /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             4950..5003
                     /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             5004..5603
                     /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    5604..6191
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6265..6498
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6570..6691
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6710..6845
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6883..6899)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6907..6923
                     /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(6931..6961)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6976..6997
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7285..7873)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8044..8904)
                     /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(8905..9009)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9483..9499
                     /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 gccaccatgg ccttcattgt agctatgggg atgattttaa tggctggaat
     2341 ctgtcctgct gtcctatgct tcccagatgg cacaaaggaa atggacattg tattccatga
     2401 acaccaagac aatgggacac aagatgacag tctcacattg gcctccgtca acactgactt
     2461 tgccttcagt ctgtacaaga agctggcttt gaagaatcaa gataaaaata ttgtcttctc
     2521 cccacttagc atctcagctg ccttggccct cgtgtccctg ggagcaaagg gcaagaccat
     2581 ggaagagatt ttagaaggcc tcaagttcaa tctcacagag acccctgaag cagacatcca
     2641 ccagggcttt ggcaacctcc tacagagtct cagccagcca gaagaccagg atcagataaa
     2701 cataggcaat gccatgttta ttgaaaagga cctgcagatc ctggcagaat tccatgagaa
     2761 gacaagggct ctgtaccaga ctgaggcctt cacagcagac ttccagcagc ctactgaggc
     2821 caaaaacctc atcaatgact atgtgagcaa tcagacccag gggatgatca agaaactgat
     2881 ctcagaactg gatgatggga cattgatggt gctggtgaat tacatctact ttaaaggcaa
     2941 atggaagata tcctttgacc cccaggacac atttgagtct gagttctact tggatgagaa
     3001 gagatctgtg aaggttccca tgatgaaaat gaagttactg actgcacgcc acttccgtga
     3061 tgaggagcta tcgtgctctg tgttggagct gaagtacaca ggaaatgcca gtgccctgct
     3121 catcctccct gaccaaggca ggatgcagca ggtggaagcc agcttacaac cagagaccct
     3181 gaggaaatgg aggaaaactt tgttttccag ccaaatagag gagctaaacc tgcccaagtt
     3241 ctccatcgct agtgactaca ggctggaaga ggatgtcctt ccagaaatgg ggattaagga
     3301 agtcttcaca gaacaagctg acctatctgg gatcacagaa gccaagaaac tgagtgtgtc
     3361 tcaggtggtc cacaaggctg tgctggatgt ggctgagaca ggcacagaag cagctgctgc
     3421 cacaggggtt attggtggca ttcgtaaggc cgtattacca gctgtgtgtt tcaacaggcc
     3481 attcctgatt gttatctatc acacaagtgc tcagagtatc ctctttatgg ccaaagtcaa
     3541 taaccccaag ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga
     3601 ctacaaggat gacgatgaca agtgagcggc cgcgaaggat ctgcgatcgc tccggtgccc
     3661 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3721 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3781 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3841 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     3901 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     3961 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     4021 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     4081 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     4141 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga
     4201 gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg
     4261 gcgtggagtt cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca
     4321 acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga
     4381 tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc
     4441 acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg
     4501 tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg
     4561 tggtgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca
     4621 acgccaccgt ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc
     4681 gcaccttcag cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact
     4741 tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc
     4801 gcgtggagga gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca
     4861 agacccccat cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca
     4921 ccgccggacc cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg
     4981 acgtggagga gaatcccggc cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5041 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     5101 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5161 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5221 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5281 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5341 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5401 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5461 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5521 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5581 tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gaaagattga
     5641 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5701 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5761 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5821 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5881 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5941 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6001 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6061 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6121 ctctgcggcc tcttccgcgt ctccgccttc gccctcagac gagtcggatc tccctttggc
     6181 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6241 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc
     6301 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6361 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6421 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6481 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     6541 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     6601 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6661 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     6721 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6781 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6841 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     6901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7021 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7081 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7201 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7261 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7321 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7381 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7441 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7501 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7621 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7681 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7741 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7801 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     9061 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     9121 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     9181 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     9241 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     9301 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     9361 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     9421 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     9481 ttgtaaaacg acggccagtg ccaagctg
//
