LOCUS       V047010                 7234 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047010
VERSION     V047010
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..2670
                     /label="Il22ra1(NM_178257)"
                     /note="Il22ra1(NM_178257)"
                     /gene="Il22ra1"
     CDS             2677..2700
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2707..2796
                     /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    2834..3058
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3104..3532
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3546..3875
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3942..4736
                     /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    4910..5031
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5080..5096)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5104..5120
                     /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(5128..5158)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5173..5194
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5482..6067)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6238..7098)
                     /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(7099..7203)
                     /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 aagacactac tgaccatcct gacggtggga
      961 tccctggccg ctcacaccac tgtggacaca tccggtctcc ttcaacacgt gaaattccag
     1021 tccagcaact ttgagaacat cttgacgtgg gatggtgggc ccgctagcac ctctgacacc
     1081 gtctacagtg tggaatataa gaaatacgga gagagaaagt ggctggccaa ggcgggctgc
     1141 cagcggatca cccagaagtt ctgcaacctg actatggaga cccgcaacca cactgagttt
     1201 tactacgcca aggtcacggc ggtcagcgcg ggaggcccac cagtcacaaa gatgactgat
     1261 cgtttcagct cgctgcagca cactaccatc aaaccgcctg atgtgacctg tatccccaaa
     1321 gtgaggtcca ttcagatgct ggtccacccc acactcacac cggtcctctc ggaagatggc
     1381 caccagctaa ccctggagga gattttccat gacctgttct accgcttaga gctccacgtc
     1441 aaccacacct accagatgca ccttgaaggc aaacagagag aatacgagtt ccttggcctg
     1501 actcccgaca cagagttcct cggctccatc acaattttga ctccgatatt gtccaaggaa
     1561 agtgccccct acgtgtgccg agtgaagacg ctgcccgatc ggacgtgggc ctactccttc
     1621 tcgggcgccg tgctcttttc catgggtttc ctcgtcggct tgctctgtta tctgggctac
     1681 aaatacatca ccaagccacc tgtacctcct aactccctga acgtccaacg tgtcctgacc
     1741 tttcaacccc tacgcttcat ccaagaacac gtactgatcc ctgtcttgga cctcagtggc
     1801 cccagcagtc tgcctcagcc catccagtac tcccaagtgg tggtgtctgg gcccagggag
     1861 cctcctggag ctgtgtggcg gcagagcctg tctgacctca cctacgtagg gcagtcagat
     1921 gtctccatcc tgcaacctac caacgtgcca gctcagcaga cactgtcccc accatcctac
     1981 gctccgaagg ctgtccctga ggtccagccc ccttcctatg cgcctcaggt agcctcggat
     2041 gccaaagctc tgttctactc accacaacag gggatgaaga ccaggcctgc cacctatgac
     2101 ccgcaggaca ttctggacag ctgccctgct tcttatgctg tgtgtgtgga agactctggc
     2161 aaagactcta ccccaggcat cctctccact cccaaatacc tcaagacaaa aggtcagctc
     2221 caggaagaca cacttgttag aagctgtctc ccaggggacc tttctctaca gaaagtcacc
     2281 tccttaggtg aaggggagac acagagacca aaatcactcc cctcacctct gggattttgc
     2341 acagacagag gacctgacct tcacacactg cgcagtgagg aaccagagac accacggtac
     2401 ctgaaggggg cgctgtctct cctgtcctct gtgcagatcg agggccaccc tgtctccctc
     2461 cctttgcacg tccattctgt ctcatgttcc ccctcagacg agggaccaag tccctggggc
     2521 ctgctggact cccttgtgtg tccaaaggat gagggtcccg cggttgagac tgaggccatg
     2581 tgccccagtg ctgcagcctc tgagctggag cagtccacag aactggactc tcttttcaaa
     2641 ggcttggccc tgactgtgca gtgggaatcc ctcgaggatt acaaggatga cgacgataag
     2701 ggatccagcg cctggtccca ccctcagttt gagaagggcg gaggctctgg cggcggaagc
     2761 ggaggatctg cttggagcca cccccagttc gaaaagtgat gagggcccgt ttaaacccgc
     2821 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2881 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2941 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3001 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3061 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3121 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3181 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3241 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3301 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3361 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3421 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3481 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3541 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3601 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3721 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3781 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3841 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3901 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3961 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4021 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4081 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4141 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4201 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4261 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4321 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4381 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4441 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4501 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4561 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4621 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4681 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4741 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4801 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4861 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4921 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4981 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5041 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5101 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5161 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5221 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5281 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5341 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5401 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5461 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5521 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5581 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5641 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5701 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5761 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5821 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5881 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5941 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6001 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6121 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6181 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6241 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6301 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6361 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6421 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6481 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6541 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6601 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6661 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6721 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6781 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6841 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6901 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6961 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7021 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7081 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7141 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7201 gcgcacattt ccccgaaaag tgccacctga cgtc
//
