LOCUS       V042124                 9318 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042124
VERSION     V042124
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2290..3408
                     /label="Ccr2(NM_009915)"
                     /note="Ccr2(NM_009915)"
                     /gene="Ccr2"
     regulatory      3415..3424
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3424..3489
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3539..3750
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3763..4031
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4066..4758
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             4759..4812
                     /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             4813..5412
                     /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    5413..6001
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6075..6308
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6380..6501
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6520..6655
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6693..6709)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6717..6733
                     /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(6741..6771)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6786..6807
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7095..7683)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7854..8714)
                     /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(8715..8819)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9293..9309
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagccacca tggaagacaa taatatgtta cctcagttca tccacggcat actatcaaca
     2341 tctcattctc tatttacacg aagtatccaa gagcttgatg aaggggccac cacaccgtat
     2401 gactacgatg atggtgagcc ttgtcataaa accagtgtga agcaaattgg agcttggatc
     2461 ctgcctccac tctactccct ggtattcatc tttggttttg tgggcaacat gttggtcatt
     2521 ataattctga taggctgtaa aaagctgaag agcatgactg atatctatct gctcaacttg
     2581 gccatctctg acctgctctt cctgctcaca ttaccattct gggctcacta tgctgcaaat
     2641 gagtgggtct ttgggaatat aatgtgtaaa gtattcacag ggctctatca cattggttat
     2701 tttggtggaa tctttttcat tatcctcctg acaattgata ggtacttggc tattgttcat
     2761 gctgtgtttg ctttaaaagc caggacagtt acctttgggg tgataacaag tgtagtcact
     2821 tgggtggtgg ctgtgtttgc ctctctacca ggaatcatat ttactaaatc caaacaagat
     2881 gatcaccatt acacctgtgg cccttatttt acacaactat ggaagaattt ccaaacaata
     2941 atgagaaata tcttgagcct gatcctgcct ctacttgtca tggtcatctg ctactcagga
     3001 attctccaca ccctgtttcg ctgtaggaat gagaagaaga ggcacagggc tgtgaggctc
     3061 atctttgcca tcatgattgt ctactttctc ttctggactc catacaatat tgttctcttc
     3121 ttgaccacct tccaggaatc cttgggaatg agtaactgtg tgattgacaa gcacttagac
     3181 caggccatgc aggtgacaga gactcttgga atgacacact gctgcattaa tcctgtcatt
     3241 tatgcctttg ttggagagaa gttccgaagg tatctctcca tatttttcag aaagcacatt
     3301 gctaaacgtc tctgcaaaca gtgcccagtt ttctataggg agacagcaga tcgagtgagc
     3361 tctacattca ctccttccac tggggagcaa gaggtctcgg ttgggttgga attcgccacc
     3421 atggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3481 gatgacaagt agggatccgc ggccgcaagg atctgcgatc gctccggtgc ccgtcagtgg
     3541 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     3601 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     3661 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     3721 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     3781 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     3841 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     3901 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     3961 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     4021 cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgag cgagctgatc
     4081 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag
     4141 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgaa gatcaaggtg
     4201 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc
     4261 agcaaagcct tcatcaacca cacccagggc atccccgact tctttaagca gtccttccct
     4321 gagggcttca catgggagag aatcaccaca tacgaagacg ggggcgtgct gaccgctacc
     4381 caggacacca gcttccagaa cggctgcatc atctacaacg tcaagatcaa cggggtgaac
     4441 ttcccatcca acggccctgt gatgcagaag aaaacacgcg gctgggaggc caacaccgag
     4501 atgctgtacc ccgctgacgg cggcctgaga ggccacagcc agatggccct gaagctcgtg
     4561 ggcgggggct acctgcactg ctccttcaag accacataca gatccaagaa acccgctaag
     4621 aacctcaaga tgcccggctt ccacttcgtg gaccacagac tggaaagaat caaggaggcc
     4681 gacaaagaga cctacgtcga gcagcacgag atggctgtgg ccaagtactg cgacctccct
     4741 agcaaactgg ggcacagaga gggcagagga agtcttctaa catgcggtga cgtggaggag
     4801 aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4861 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4921 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4981 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5041 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5101 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5161 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5221 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5281 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5341 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5401 cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5461 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5521 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5581 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5641 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5701 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5761 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5821 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5881 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5941 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6001 cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6061 agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc
     6121 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6181 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6241 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6301 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6361 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     6421 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6481 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     6541 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6601 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
     6661 ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6721 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6781 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6841 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6901 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6961 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7501 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7741 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7801 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7861 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7921 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7981 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8041 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8101 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8161 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8221 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8281 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8341 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8401 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8461 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8521 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8581 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8641 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8701 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8821 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8881 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     8941 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     9001 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     9061 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     9121 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     9181 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     9241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     9301 acggccagtg ccaagctg
//
