LOCUS       V028012                 9094 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028012
VERSION     V028012
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..3120
                     /label="Mettl1(NM_010792)"
                     /note="Mettl1(NM_010792)"
                     /gene="Mettl1"
     CDS             3127..3192
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3236..3447
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3460..3728
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3757..3766
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3787..4449
                     /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             4519..4572
                     /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             4573..5172
                     /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    5173..5761
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5835..6068
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6140..6261
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6301..6436
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6469..6485)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6493..6509
                     /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(6517..6547)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6562..6583
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6871..7459)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7630..8490)
                     /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(8491..8595)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9069..9085
                     /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 gccaccatga tggcgggagc cgaagccccc
     2341 cagccgcaga agcgctacta ccggcagcgc gcccactcca accccatggc agaccacaca
     2401 ctgcgctacc ctgtgaagcc agaggaaatg gactggtctg agctttaccc agagttcttt
     2461 gctccgctta ttcaaaataa gagccatgat gatccaaagg atgagaaaga aaagcactct
     2521 ggggcccaag tggagtttgc agacataggc tgtggctatg gtggcttgtt agtggcactg
     2581 tcaccgctct tcccagatac cctgattctg ggtctggaga ttcgggtgaa ggtgtcggac
     2641 tatgtgcagg acaggattcg ggccctccgt gcagctcccg ggggcggatt ccagaacatc
     2701 gcctgtctcc gaagtaacgc catgaaacac cttcctaatt tcttccgcaa gggccagctg
     2761 gcaaagatgt tcttcctctt cccggaccca cactttaagc gaacgaagca taaatggaga
     2821 atcatcagcc ctacgcttct ggcagagtat gcctacgtgc tgagagtcgg gggcctggtg
     2881 tacaccgtca ccgacgtgcc ggagctgcat gagtggatgt gcacccactt tgaagaacac
     2941 ccactgtttg agcgtgtgcc tcttgaagag ctaagtgaag accccattgt ggaacatcta
     3001 ggcagttcaa ctgaggaagg aaagaaagtt ctacgcaatg ggggaaagaa tttcccagcc
     3061 gtcttccgaa gaatacagga tcccctcctc caggcagtga cccccaaccc caccctgcct
     3121 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     3181 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     3241 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     3301 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     3361 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     3421 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     3481 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     3541 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     3601 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     3661 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     3721 tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag cgacgagagc
     3781 ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc
     3841 gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa caagatgaag
     3901 agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc
     3961 ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac
     4021 aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg
     4081 agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc
     4141 ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg
     4201 gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc
     4261 ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc
     4321 atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag
     4381 ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc
     4441 gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac cgccggaccc
     4501 ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag
     4561 aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4621 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4681 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4741 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4801 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4861 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4921 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4981 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5041 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5101 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5161 cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5221 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5281 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5341 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5401 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5461 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5521 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5581 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5641 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5701 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5761 cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5821 agaaaagggg ggactggaag ggctaattca ctcccaacga agataagatc tgctttttgc
     5881 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5941 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6001 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6061 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6121 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     6181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6241 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     6301 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6361 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6421 ggcttttttg gaggcctaga cttttgcaga gaccaaattc gtaatcatgt catagctgtt
     6481 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6541 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8641 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8701 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8761 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8821 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     8881 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     8941 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9001 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9061 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
