LOCUS       V027042                 9541 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027042
VERSION     V027042
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..3867
                     /label="ERLIN1(NM_001100626)"
                     /note="ERLIN1(NM_001100626)"
                     /gene="ERLIN1"
     CDS             3904..4623
                     /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        4673..5002
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5012..5611
                     /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    5678..6266
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6353..6586
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6658..6779
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6819..6954
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6975..6993)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7003..7019)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      7161..7616
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7642..7746
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7747..8607
                     /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      8778..9366
                     /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 accatgaata tgactcaagc ccgggttctg gtggctgcag tggtggggtt ggtggctgtc
     2881 ctgctctacg cctccatcca caagattgag gagggccatc tggctgtgta ctacagggga
     2941 ggagctttac taactagccc cagtggacca ggctatcata tcatgttgcc tttcattact
     3001 acgttcagat ctgtgcagac aacactacaa actgatgaag ttaaaaatgt gccttgtgga
     3061 acaagtggtg gggtcatgat ctatattgac cgaatagaag tggttaatat gttggctcct
     3121 tatgcagtgt ttgatatcgt gaggaactat actgcagatt atgacaagac cttaatcttc
     3181 aataaaatcc accatgagct gaaccagttc tgcagtgccc acacacttca ggaagtttac
     3241 attgaattgt ttgatcaaat agatgaaaac ctgaagcaag ctctgcagaa agacttaaac
     3301 ctcatggccc caggtctcac tatacaggct gtgcgtgtta caaaacccaa aatcccagaa
     3361 gccataagaa gaaattttga gttaatggag gctgagaaga caaaactcct tatagctgca
     3421 cagaaacaaa aggttgtgga aaaagaagct gagacagaga ggaaaaaggc agttatagaa
     3481 gcagagaaga ttgcacaagt ggcaaaaatt cggtttcagc agaaagtgat ggaaaaagaa
     3541 actgaaaagc gcatttctga aatcgaagat gctgcattcc tggcccgaga gaaagcgaaa
     3601 gcagatgctg aatattatgc tgcacacaaa tatgccacct caaacaagca caagttgacc
     3661 ccggaatatc tggagctcaa aaagtaccag gccattgctt ctaacagtaa gatctatttt
     3721 ggcagcaaca tccctaacat gttcgtggac tcctcatgtg ctttgaaata ttcagatatt
     3781 aggactggaa gagaaagctc actcccctct aaggaggctc ttgaaccctc tggagagaac
     3841 gtcatccaaa acaaagagag cacaggtctc gagggcggtg gaggcagtgg tggaggcggt
     3901 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3961 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4021 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4081 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4141 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4201 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4261 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4321 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4381 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4441 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4501 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4561 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4621 taacgggatc cggactagtt aacgcgtcga ccagtggtcg accctgtgga atgtgtgtca
     4681 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4741 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4801 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4861 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4921 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4981 tggaggccta ggcttttgca aaaagcttac catgaccgag tacaagccca cggtgcgcct
     5041 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     5101 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     5161 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5221 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5281 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5341 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5401 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5461 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5521 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5581 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     5641 cgaaaggagc gcacgacccc atgcatcgcc agtcgacaat caacctctgg attacaaaat
     5701 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5761 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5821 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5881 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5941 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6001 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6061 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     6121 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6181 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6241 gatctccctt tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg
     6301 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     6361 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     6421 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6481 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6541 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     6601 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     6661 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6721 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6781 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga
     6961 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     7021 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     7081 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     7141 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     7201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     7261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     7321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     7381 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7441 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     7501 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     7561 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     7621 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7681 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7741 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7801 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7861 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7921 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7981 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     8041 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     8101 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     8161 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     8221 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     8281 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     8341 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     8401 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8461 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8521 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8581 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8641 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8701 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8761 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8821 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8881 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8941 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     9001 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     9061 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     9121 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     9181 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     9241 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     9301 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     9361 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     9421 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     9481 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9541 g
//
