LOCUS       V042071                 7213 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042071
VERSION     V042071
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             928..2649
                     /label="IL22RA1(NM_021258)"
                     /note="IL22RA1(NM_021258)"
                     /gene="IL22RA1"
     CDS             2656..2679
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2686..2775
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    2813..3037
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3083..3511
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3525..3854
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3921..4715
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4889..5010
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5059..5075)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5083..5099
                     /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"
     promoter        complement(5107..5137)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5152..5173
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5461..6046)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6217..7077)
                     /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"
     promoter        complement(7078..7182)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctgaatt cgccaccatg aggacgctgc tgaccatctt gactgtggga
      961 tccctggctg ctcacgcccc tgaggacccc tcggatctgc tccagcacgt gaaattccag
     1021 tccagcaact ttgaaaacat cctgacgtgg gacagcgggc cggagggcac cccagacacg
     1081 gtctacagca tcgagtataa gacgtacgga gagagggact gggtggcaaa gaagggctgt
     1141 cagcggatca cccggaagtc ctgcaacctg acggtggaga cgggcaacct cacggagctc
     1201 tactatgcca gggtcaccgc tgtcagtgcg ggaggccggt cagccaccaa gatgactgac
     1261 aggttcagct ctctgcagca cactaccctc aagccacctg atgtgacctg tatctccaaa
     1321 gtgagatcga ttcagatgat tgttcatcct acccccacgc caatccgtgc aggcgatggc
     1381 caccggctaa ccctggaaga catcttccat gacctgttct accacttaga gctccaggtc
     1441 aaccgcacct accaaatgca ccttggaggg aagcagagag aatatgagtt cttcggcctg
     1501 acccctgaca cagagttcct tggcaccatc atgatttgcg ttcccacctg ggccaaggag
     1561 agtgccccct acatgtgccg agtgaagaca ctgccagacc ggacatggac ctactccttc
     1621 tccggagcct tcctgttctc catgggcttc ctcgtcgcag tactctgcta cctgagctac
     1681 agatatgtca ccaagccgcc tgcacctccc aactccctga acgtccagcg agtcctgact
     1741 ttccagccgc tgcgcttcat ccaggagcac gtcctgatcc ctgtctttga cctcagcggc
     1801 cccagcagtc tggcccagcc tgtccagtac tcccagatca gggtgtctgg acccagggag
     1861 cccgcaggag ctccacagcg gcatagcctg tccgagatca cctacttagg gcagccagac
     1921 atctccatcc tccagccctc caacgtgcca cctccccaga tcctctcccc actgtcctat
     1981 gccccaaacg ctgcccctga ggtcgggccc ccatcctatg cacctcaggt gacccccgaa
     2041 gctcaattcc cattctacgc cccacaggcc atctctaagg tccagccttc ctcctatgcc
     2101 cctcaagcca ctccggacag ctggcctccc tcctatgggg tatgcatgga aggttctggc
     2161 aaagactccc ccactgggac actttctagt cctaaacacc ttaggcctaa aggtcagctt
     2221 cagaaagagc caccagctgg aagctgcatg ttaggtggcc tttctctgca ggaggtgacc
     2281 tccttggcta tggaggaatc ccaagaagca aaatcattgc accagcccct ggggatttgc
     2341 acagacagaa catctgaccc aaatgtgcta cacagtgggg aggaagggac accacagtac
     2401 ctaaagggcc agctccccct cctctcctca gtccagatcg agggccaccc catgtccctc
     2461 cctttgcaac ctccttcccg tccatgttcc ccctcggacc aaggtccaag tccctggggc
     2521 ctgctggagt cccttgtgtg tcccaaggat gaagccaaga gcccagcccc tgagacctca
     2581 gacctggagc agcccacaga actggattct cttttcagag gcctggccct gactgtgcag
     2641 tgggagtccc tcgaggatta caaggatgac gacgataagg gatccagcgc ctggtcccac
     2701 cctcagtttg agaagggcgg aggctctggc ggcggaagcg gaggatctgc ttggagccac
     2761 ccccagttcg aaaagtgatg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2821 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2881 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2941 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3001 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3061 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3121 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3181 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3241 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3301 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3361 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3421 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3481 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3541 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3601 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3661 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3721 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3781 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3841 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3901 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3961 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4021 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4081 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4141 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4201 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4261 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4321 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4381 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4441 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4501 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4561 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4621 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4681 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4741 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4801 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4861 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4921 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4981 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5041 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7081 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7141 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7201 gccacctgac gtc
//
