LOCUS       V019771                 9007 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019771
VERSION     V019771
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2058..2175
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        2227..2530
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2531..2734
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2854..3570
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3586..4137
                     /label="TNFAIP8L2(NM_024575)"
                     /note="TNFAIP8L2(NM_024575)"
                     /gene="TNFAIP8L2"
     CDS             4156..4245
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4263..4763
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4784..5383
                     /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    5397..5985
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6060..6293
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        6498..6602
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6603..7463
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      7634..8222
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    8633..8654
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        8669..8699
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8707..8723
                     /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"
     primer_bind     8731..8747
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccac gcgtcgaagc
     2821 ccgggcggat cgggtttaaa cggatccgcc accatggtga gcaagggcga ggagctgttc
     2881 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2941 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     3001 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     3061 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3121 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3181 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3241 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3301 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3361 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3421 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     3481 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3541 atcactctcg gcatggacga gctgtacaag ggaggagggg gaagcatgga gtccttcagc
     3601 tcaaagagcc tggcactgca agcagagaag aagctactga gtaagatggc gggtcgctct
     3661 gtggctcatc tcttcataga tgagacaagc agtgaggtgc tagatgagct ctaccgtgtg
     3721 tccaaggagt acacgcacag ccggccccag gcccagcgcg tgatcaagga cctgatcaaa
     3781 gtggccatca aggtggctgt gctgcaccgc aatggctcct ttggccccag tgagctggcc
     3841 ctggctaccc gctttcgcca gaagctgcgg cagggtgcca tgacggcact tagctttggt
     3901 gaggtagact tcaccttcga ggctgctgtt ctggctggcc tgctgaccga gtgccgggat
     3961 gtgctgctag agttggtgga acaccacctc acgcccaagt cacatggccg catccgccac
     4021 gtgtttgatc acttctctga cccaggtctg ctcacggccc tctatgggcc tgacttcact
     4081 cagcaccttg gcaagatctg tgacggactc aggaagctgc tagacgaagg gaagctcctc
     4141 gagtctagag ggcccgagca gaaactcatc tctgaagaag atctggagca gaaactcatc
     4201 tctgaagaag atctggagca gaaactcatc tctgaagaag atctgtagtc gagaattcta
     4261 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     4321 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     4381 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     4441 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     4501 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     4561 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     4621 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg ccccgaaggt
     4681 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     4741 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc
     4801 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4861 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4921 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4981 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5041 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5101 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5161 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5221 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5281 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5341 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaatcgata gatcctaatc
     5401 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5461 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5521 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5581 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5641 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5701 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5761 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     5821 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5881 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5941 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgag atcctttaag
     6001 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     6061 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     6121 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6181 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6241 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     6301 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     6361 gaggcccggg ttaattaagg aaagggctag atcattcttg aagacgaaag ggcctcgtga
     6421 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6481 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6841 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6901 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6961 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7021 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7081 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7141 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7201 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7261 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7321 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7381 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7441 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7501 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7561 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7621 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7681 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7741 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7801 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7861 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7921 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7981 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8041 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8101 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8161 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8221 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8281 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8341 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8401 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagca
     8461 agctcatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     8521 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     8581 cgtggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     8641 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     8701 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt
     8761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     8821 tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc
     8881 ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg
     8941 attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact
     9001 tagtagt
//
