LOCUS       V028800                 9985 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028800
VERSION     V028800
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        9..235
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             236..416
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    463..588
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1081..1314
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1499..1543
                     /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    1810..1926
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2130..3312
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3335..4768
                     /label="SLC16A7(NM_001270623)"
                     /note="SLC16A7(NM_001270623)"
                     /gene="SLC16A7"
     misc_feature    4817..5390
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5424..5822
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5837..6425
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6512..6745
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6817..6938
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6978..7113
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(7134..7152)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7162..7178)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      7320..7775
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7801..7905
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7906..8766
                     /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      8937..9525
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9813..9834
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9849..9879
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9887..9903
                     /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     9911..9927
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9948..9966
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
       61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
      121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
      181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
      541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
      601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
      661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
      721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
      781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
      841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
      901 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
      961 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1021 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1081 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1141 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1201 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1261 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1321 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1381 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     1441 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     1501 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     1561 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     1621 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     1681 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     1741 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     1801 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     1861 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa
     1921 aattttatcg atgcctcccc gtcaccaccc cccccaaccc gccccgaccg gagctgagag
     1981 taattcatac aaaaggactc gcccctgcct tggggaatcc cagggaccgt cgttaaactc
     2041 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc
     2101 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca
     2161 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     2221 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2281 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2341 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     2401 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt
     2461 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     2521 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     2581 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     2641 ttttgatgat atcctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     2701 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     2761 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     2821 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     2881 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     2941 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     3001 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     3061 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     3121 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     3181 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     3241 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     3301 caggtgtcgt gaaaactacc cctctagagc caccatgcca ccaatgccaa gtgccccacc
     3361 tgtgcatcca cctccagatg gaggatgggg ttggattgtg gttggagcag cttttatctc
     3421 cattggattt tcctatgcat tccccaaagc tgtcaccgta ttcttcaaag aaattcagca
     3481 aatattccac actacctaca gtgaaatagc atggatttca tccattatgc tggctgttat
     3541 gtacgcagga ggtcctgtaa gtagtgtttt ggtgaataaa tacggcagcc ggccggtggt
     3601 gatagcagga ggcttattat gctgtcttgg aatggtgttg gcctccttta gtagcagcgt
     3661 ggtacagctg tacctcacta tgggattcat tacaggttta ggtttagcct tcaacctgca
     3721 acccgcctta accataattg gcaaatactt ctataggaag cgacccatgg caaatggatt
     3781 ggccatggca ggaagtcctg ttttcttaag ttcattggct cctttcaatc agtacctttt
     3841 taatactttt ggctggaaag gaagcttcct gattttggga agtctacttt tgaatgcctg
     3901 tgtggctggt tccctcatga gaccccttgg acccaatcaa accacttcta agtctaaaaa
     3961 taagactggc aaaacagaag atgattcaag cccaaagaaa atcaaaacga agaaatcaac
     4021 ttgggaaaaa gttaataagt atttagattt ctcccttttt aagcatagag gatttctgat
     4081 atatctgtct ggaaatgtca ttatgttcct aggttttttt gcccccatta tattcttggc
     4141 tccatatgct aaagaccaag gaattgatga gtactcggca gcttttctgc tatctgttat
     4201 ggctttcgtt gatatgtttg ctaggccttc tgtaggatta attgcaaact ccaaatatat
     4261 tcgacctcga attcagtact tcttcagttt tgcaatcatg ttcaatggag tgtgtcacct
     4321 cttgtgccca ctggcacagg actacacaag cctggtatta tatgctgtat tttttggcct
     4381 tggatttggg agtgttagca gtgttctctt tgaaactctc atggacctcg tgggtgcacc
     4441 aagattttcc agtgccgtcg gacttgtcac aattgtggag tgtggcccag ttcttcttgg
     4501 ccctcctctt gcaggtaaat tggtggattt aactggagaa tataaataca tgtacatgtc
     4561 ctgtggggct attgtggtag cagcaagcgt gtggctgctc attggcaatg ctatcaacta
     4621 tagattgctt gcaaaggaaa ggaaggagga aaatgcaagg cagaagacca gagaatctga
     4681 acccttgagc aaatctaaac attcggaaga tgttaacgtc aaagtttcaa atgcacagag
     4741 tgtaacctca gaaagagaaa ctaacattta aggatccacg cgtctccggc ctagggataa
     4801 cagggtaatc cgctagcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     4861 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4921 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4981 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     5041 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     5101 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     5161 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     5221 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     5281 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     5341 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     5401 ctagagatcc accggtcgcc accatggcca agcctttgtc tcaagaagaa tccaccctca
     5461 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     5521 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     5581 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     5641 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     5701 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     5761 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     5821 aagcggccgc gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5881 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5941 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6001 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6061 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6121 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6181 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6241 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6301 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6361 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6421 cccgcctgga attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat
     6481 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     6541 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6601 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6661 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6721 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6781 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6841 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6901 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6961 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     7021 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     7081 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     7141 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     7201 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     7261 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     7321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     7381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     7441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     7561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     7681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     7741 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     7801 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7861 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7921 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7981 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     8041 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     8101 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     8161 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     8221 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     8281 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     8341 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     8401 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     8461 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     8521 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     8581 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     8641 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8701 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8761 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8821 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8881 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8941 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     9001 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     9061 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     9121 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     9181 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     9241 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     9301 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     9361 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     9421 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     9481 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     9541 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     9601 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     9661 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     9721 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     9781 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     9841 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     9901 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     9961 taaagggaac aaaagctgga gctgc
//
