LOCUS       V033798                 9525 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033798
VERSION     V033798
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             1494..1538
                     /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    1801..1916
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        1974..2277
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2278..2481
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2594..3595
                     /label="Mdh1(NM_008618)"
                     /note="Mdh1(NM_008618)"
                     /gene="Mdh1"
     CDS             3620..3691
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     promoter        3745..3956
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3969..4237
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4265..4957
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Zoanthus green fluorescent protein"
                     /transl_table=1
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
                     /label="ZsGreen1"
     CDS             4967..5020
                     /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             5021..5620
                     /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    5621..6208
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6282..6515
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6587..6708
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6727..6862
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6900..6916)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6924..6940
                     /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(6948..6978)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6993..7014
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7302..7890)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8061..8921)
                     /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(8922..9026)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9500..9516
                     /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 gggcgcagcg 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 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc
     1801 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     1861 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     1921 gtttattaca gggacagcag agatccagtt tatcgataag cttgggagtt ccgcgttaca
     1981 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2041 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2281 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2341 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2401 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2461 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     2521 cgctgttttg acctccatag aagacaccga ctctactaga ggatctattt ccggtgaatt
     2581 cctcgaggcc accatgtctg aaccaatcag agtccttgtg actggagcag ctggtcaaat
     2641 tgcatattca ctgttgtaca gtattggaaa tggatctgtc tttgggaaag accagcccat
     2701 cattcttgtg ctgttggaca tcacccccat gatgggtgtt ctggacggtg tcctgatgga
     2761 actgcaagac tgtgcccttc cccttctgca ggatgtcatt gcaacggaca aagaagagat
     2821 tgccttcaaa gacctggatg tggctgtcct agtgggctcc atgccaagaa gggaaggcat
     2881 ggagaggaag gacctactga aagccaatgt gaaaatcttc aaatcccagg gcacagcctt
     2941 ggagaaatac gccaagaaat cagttaaggt cattgttgtg ggaaacccag ccaatacgaa
     3001 ctgcctgaca gcctccaagt cagcgccatc gatccccaag gagaatttca gttgcctgac
     3061 tcgcttggac cacaaccgag caaaatctca aattgctctt aaactcggtg taaccgctga
     3121 tgatgtaaag aatgtcatta tctggggaaa tcattcatcg acccagtatc cagatgtcaa
     3181 tcatgccaag gtgaaactgc aaggaaagga agtcggtgtg tatgaagccc tgaaagacga
     3241 cagctggctg aagggagagt tcatcacgac tgtgcaacag cgtggtgctg ctgtcatcaa
     3301 ggctcggaag ctgtccagtg caatgtctgc tgcgaaagcc atcgcagacc acatcagaga
     3361 catctggttt ggaaccccag agggagagtt cgtgtcgatg ggtgttatct ctgatggcaa
     3421 ctcctatggt gtccctgatg acctgctcta ctcattccct gtcgtgatca agaataagac
     3481 ctggaagttt gttgaaggcc tccccattaa tgacttctcc cgtgaaaaga tggacctgac
     3541 agcaaaggag ctgaccgagg aaaaggagac cgcttttgag tttctctcct ctgcgggatc
     3601 ctcggtacca agcttaagtg actacaagga tgacgatgac aaggattaca aagacgacga
     3661 tgataaggac tataaggatg atgacgacaa ataaagatcc atcgatacta gtaaggatct
     3721 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     3781 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     3841 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     3901 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     3961 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     4021 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     4081 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     4141 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     4201 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctactc
     4261 tagcatggcc cagtccaagc acggcctgac caaggagatg accatgaagt accgcatgga
     4321 gggctgcgtg gacggccaca agttcgtgat caccggcgag ggcatcggct accccttcaa
     4381 gggcaagcag gccatcaacc tgtgcgtggt ggagggcggc cccttgccct tcgccgagga
     4441 catcttgtcc gccgccttca tgtacggcaa ccgcgtgttc accgagtacc cccaggacat
     4501 cgtcgactac ttcaagaact cctgccccgc cggctacacc tgggaccgct ccttcctgtt
     4561 cgaggacggc gccgtgtgca tctgcaacgc cgacatcacc gtgagcgtgg aggagaactg
     4621 catgtaccac gagtccaagt tctacggcgt gaacttcccc gccgacggcc ccgtgatgaa
     4681 gaagatgacc gacaactggg agccctcctg cgagaagatc atccccgtgc ccaagcaggg
     4741 catcttgaag ggcgacgtga gcatgtacct gctgctgaag gacggtggcc gcttgcgctg
     4801 ccagttcgac accgtgtaca aggccaagtc cgtgccccgc aagatgcccg actggcactt
     4861 catccagcac aagctgaccc gcgaggaccg cagcgacgcc aagaaccaga agtggcacct
     4921 gaccgagcac gccatcgcct ccggctccgc cttgcccgcg gccgctgagg gcagaggaag
     4981 tcttctaaca tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac
     5041 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     5101 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     5161 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5221 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5281 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5341 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5401 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5461 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5521 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5581 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa
     5641 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5701 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5761 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5821 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5881 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5941 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6001 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     6061 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6121 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc cgccttcgcc ctcagacgag
     6181 tcggatctcc ctttggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc
     6241 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     6301 ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     6361 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6421 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6481 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6541 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6601 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6661 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     6721 ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6781 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6841 tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg
     6901 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6961 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7021 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7741 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7921 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7981 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8041 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8101 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8161 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8221 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8281 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8341 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8401 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8461 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8521 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8581 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8641 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8701 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8761 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8821 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8881 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     9061 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     9121 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     9181 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     9241 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     9301 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     9361 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     9421 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     9481 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
