LOCUS       V026674                 9553 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026674
VERSION     V026674
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     protein_bind    113..134
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        149..179
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    187..203
                     /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     211..227
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        248..266
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        294..520
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             521..701
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    748..873
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1366..1599
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1784..1828
                     /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    2095..2211
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        2234..2537
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2538..2741
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2824..3879
                     /label="CCR5(NM_000579)"
                     /note="CCR5(NM_000579)"
                     /gene="CCR5"
     CDS             3916..4635
                     /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"
     promoter        4685..5014
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5024..5623
                     /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    5690..6278
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6365..6598
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6670..6791
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6831..6966
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6987..7005)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7015..7031)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      7173..7628
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7654..7758
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7759..8619
                     /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"
     rep_origin      8790..9378
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
       61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      241 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
      301 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
      361 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
      421 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
      481 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
      541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
      721 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
      781 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
      841 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
      901 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
      961 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     1021 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     1081 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     1141 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     1201 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     1261 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     1321 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     1381 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     1441 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     1501 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     1561 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     1621 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     1681 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     1741 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     1801 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     1861 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     1921 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     1981 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     2041 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa
     2101 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2161 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag
     2221 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2521 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2581 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2641 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2701 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     2761 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagagcc
     2821 accatggatt atcaagtgtc aagtccaatc tatgacatca attattatac atcggagccc
     2881 tgccaaaaaa tcaatgtgaa gcaaatcgca gcccgcctcc tgcctccgct ctactcactg
     2941 gtgttcatct ttggttttgt gggcaacatg ctggtcatcc tcatcctgat aaactgcaaa
     3001 aggctgaaga gcatgactga catctacctg ctcaacctgg ccatctctga cctgtttttc
     3061 cttcttactg tccccttctg ggctcactat gctgccgccc agtgggactt tggaaataca
     3121 atgtgtcaac tcttgacagg gctctatttt ataggcttct tctctggaat cttcttcatc
     3181 atcctcctga caatcgatag gtacctggct gtcgtccatg ctgtgtttgc tttaaaagcc
     3241 aggacggtca cctttggggt ggtgacaagt gtgatcactt gggtggtggc tgtgtttgcg
     3301 tctctcccag gaatcatctt taccagatct caaaaagaag gtcttcatta cacctgcagc
     3361 tctcattttc catacagtca gtatcaattc tggaagaatt tccagacatt aaagatagtc
     3421 atcttggggc tggtcctgcc gctgcttgtc atggtcatct gctactcggg aatcctaaaa
     3481 actctgcttc ggtgtcgaaa tgagaagaag aggcacaggg ctgtgaggct tatcttcacc
     3541 atcatgattg tttattttct cttctgggct ccctacaaca ttgtccttct cctgaacacc
     3601 ttccaggaat tctttggcct gaataattgc agtagctcta acaggttgga ccaagctatg
     3661 caggtgacag agactcttgg gatgacgcac tgctgcatca accccatcat ctatgccttt
     3721 gtcggggaga agttcagaaa ctacctctta gtcttcttcc aaaagcacat tgccaaacgc
     3781 ttctgcaaat gctgttctat tttccagcaa gaggctcccg agcgagcaag ctcagtttac
     3841 acccgatcca ctggggagca ggaaatatct gtgggcttgc tcgagggcgg tggaggcagt
     3901 ggtggaggcg gtagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3961 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4021 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4081 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     4141 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4201 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4261 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4321 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4381 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4441 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4501 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4561 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4621 gagctgtaca agtaacggga tccggactag ttaacgcgtc gaccagtggt cgaccctgtg
     4681 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4741 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4801 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4861 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4921 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4981 aggaggcttt tttggaggcc taggcttttg caaaaagctt accatgaccg agtacaagcc
     5041 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     5101 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     5161 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5221 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5281 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5341 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5401 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5461 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5521 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5581 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     5641 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg ccagtcgaca atcaacctct
     5701 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5761 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5821 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5881 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5941 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     6001 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     6061 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     6121 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     6181 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6241 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacct
     6301 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6361 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     6421 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6481 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6541 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6601 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6661 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6721 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6781 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     6841 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6901 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6961 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     7021 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     7081 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     7141 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     7201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     7261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7381 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     7441 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     7501 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     7561 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     7621 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7681 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7741 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7801 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7861 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7921 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7981 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     8041 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     8101 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     8161 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     8221 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     8281 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     8341 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     8401 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8461 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8521 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8581 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8641 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8701 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     9001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     9061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     9121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     9181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     9241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     9301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     9361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     9421 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     9481 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     9541 gtgagcgagg aag
//
