LOCUS       V041504                 8117 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041504
VERSION     V041504
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             2862..3284
                     /label="STUM(NM_001003665)"
                     /note="STUM(NM_001003665)"
                     /gene="STUM"
     CDS             3285..3356
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     promoter        3374..3873
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             3894..4493
                     /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    4507..5095
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5170..5403
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        5608..5712
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5713..6573
                     /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      6744..7332
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7743..7764
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7779..7809
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7817..7833
                     /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     7841..7857
                     /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 gtgaccgtca gatcgcctgg
     2761 agacgccatc cacgctgttt tgacctccat agaagacacc ggcggccacg cgtcgaagcc
     2821 cgggcggatc gggtttaaac ggatccggag gtaccgccac catggagccc tcgcacaaag
     2881 acgccgagac ggcggcggcg gcggcggcgg tggcggcggc ggacccccgg ggggcgtcct
     2941 cgtccagcgg ggtggtggtg caggtccgcg agaagaaggg ccccctgcgc gccgccatcc
     3001 cctacatgcc cttccccgtg gccgtcatct gcctcttcct caacactttc gtgccgggac
     3061 tggggacatt cgtctcggcc ttcactgtgc tgtgcggggc ccgcaccgac ctcccggaca
     3121 ggcatgtgtg ctgcgtcttc tggctgaaca ttgcagcagc cctcatccaa atcctcactg
     3181 ccatcgtcat ggtgggctgg atcatgagca tcttctgggg catggacatg gtcatccttg
     3241 ccatttccca aggctacaag gagcagggca tcccacagca gctggattac aaggatgacg
     3301 acgataagga ttacaaggat gacgacgata aggattacaa ggatgacgac gataagtagt
     3361 cgagaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
     3421 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
     3481 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
     3541 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
     3601 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
     3661 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
     3721 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
     3781 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
     3841 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg
     3901 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     3961 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4021 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4081 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4141 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4201 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4261 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4321 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     4381 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4441 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaatcgata
     4501 gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4561 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4621 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4681 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4741 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4801 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4861 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4921 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4981 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5041 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgag
     5101 atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5161 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     5221 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5281 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5341 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5401 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     5461 cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg aagacgaaag
     5521 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5941 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6241 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7501 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7561 taatgcagca agctcatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7621 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     7681 aaaagctccc cgtggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7741 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7801 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg
     7861 attacgaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     7921 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     7981 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     8041 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     8101 actacaaact tagtagt
//
