LOCUS       V032801                 9694 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032801
VERSION     V032801
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..3841
                     /label="Cyp1a2(NM_009993)"
                     /note="Cyp1a2(NM_009993)"
                     /gene="Cyp1a2"
     promoter        3892..4103
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4116..4384
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4419..5126
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             5136..5189
                     /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             5190..5789
                     /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    5790..6377
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6451..6684
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6756..6877
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6896..7031
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7069..7085)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7093..7109
                     /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(7117..7147)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7162..7183
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7471..8059)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8230..9090)
                     /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(9091..9195)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9669..9685
                     /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 ccatggcgtt ctcccagtac atctccttag ccccagagct
     2341 gctactggcc actgccatct tctgtttagt gttctggatg gtcagagcct caaggaccca
     2401 ggttcccaaa ggcctgaaga atccacccgg accctggggc ttgcccttca ttgggcacat
     2461 gctgactgtg gggaagaacc cacacctgtc actgacacgg ctgagtcagc agtatgggga
     2521 cgtgctgcag atccgcatcg gctccactcc tgtggtggtg ctgagcggcc tgaacaccat
     2581 caagcaggcc ctggtgaggc agggagatga cttcaagggc cgaccagacc tctacagctt
     2641 cacacttatc actaacggca agagcatgac tttcaaccca gactctggac ccgtgtgggc
     2701 tgcccgccgg cgcctggccc aggatgccct gaagagcttc tccatagcct cggacccgac
     2761 gtcagcatcc tcttgctatt tggaggagca cgtgagcaag gaggctaacc atctcgtcag
     2821 caagcttcag aaggcgatgg cagaggttgg ccacttcgaa ccagtcagcc aggtggtgga
     2881 atcggtggct aacgtcattg gtgccatgtg ctttgggaag aacttccccc ggaagagcga
     2941 ggagatgctg aacatcgtga ataacagcaa ggactttgtg gagaatgtca cctcagggaa
     3001 tgcagtggac ttcttcccgg tcctgcgcta cctgcccaac ccggccctca agaggtttaa
     3061 gaccttcaat gataacttcg tgctgtttct gcagaaaact gtccaggagc actaccaaga
     3121 cttcaacaag aacagtatcc aagacatcac aagtgccctg ttcaagcaca gcgagaacta
     3181 caaagacaat ggcggtctca tccccgagga gaagattgtc aacattgtca atgacatctt
     3241 tggagctggc tttgacacag tcaccacagc catcacctgg agcattttgc tacttgtgac
     3301 atggcctaac gtgcagagga agatccatga ggagctggac acggtggttg gcagggatcg
     3361 gcaaccacgg ctttctgacc gtccccagct gccatatcta gaggccttca tcctggagat
     3421 ctaccgatac acatcctttg tccccttcac catcccccac agcacaacga gggacacctc
     3481 actgaatggc ttccacattc ccaaggagcg ctgtatctac ataaaccagt ggcaggtcaa
     3541 ccatgatgag aagcagtgga aagacccctt tgtgttccgc ccagagcggt ttcttaccaa
     3601 taacaactcg gccatcgaca agacccagag cgagaaggtg atgctcttcg gcttgggaaa
     3661 gcgccggtgc attggggaga tcccggccaa gtgggaagtc ttcctcttct tagccatcct
     3721 gctgcagcat ctggagttta gtgtgccacc gggtgtgaag gtggacctga cacccaacta
     3781 tgggttgacc atgaagcccg ggacctgtga acacgtccag gcatggccac gcttttccaa
     3841 gtgaggatcc gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc
     3901 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct
     3961 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     4021 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     4081 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg
     4141 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg
     4201 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac
     4261 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt
     4321 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca
     4381 gatccaagct gtgaccggcg cctacgctag acgccaccat ggtgagcaag ggcgaggagg
     4441 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     4501 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     4561 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     4621 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     4681 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     4741 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     4801 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     4861 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     4921 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     4981 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     5041 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     5101 ccggcggcat ggacgagctg tacaagggat ctcgcgaggg cagaggaagt cttctaacat
     5161 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg
     5221 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5281 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5341 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5401 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5461 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5521 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5581 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5641 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5701 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5761 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa
     5821 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5881 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5941 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6001 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6061 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6121 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6181 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6241 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6301 tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc
     6361 tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6421 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6481 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6541 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6601 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6661 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6721 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6781 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6841 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6901 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6961 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     7021 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     7081 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7141 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7261 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     9241 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     9301 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     9361 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     9421 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9481 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9541 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9601 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9661 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
