LOCUS       V047002                 7078 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047002
VERSION     V047002
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..2514
                     /label="Il21r(NM_021887)"
                     /note="Il21r(NM_021887)"
                     /gene="Il21r"
     CDS             2521..2544
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2551..2640
                     /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    2678..2902
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2948..3376
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3390..3719
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3786..4580
                     /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    4754..4875
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4924..4940)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4948..4964
                     /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(4972..5002)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5017..5038
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5326..5911)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6082..6942)
                     /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(6943..7047)
                     /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 ccccggggcc cagtggctgc cttactcctg
      961 ctgattctcc atggagcttg gagctgcctg gacctcactt gctacactga ctacctctgg
     1021 accatcacct gtgtcctgga gacacggagc cccaacccca gcatactcag tctcacctgg
     1081 caagatgaat atgaggaact tcaggaccaa gagaccttct gcagcctaca caggtctggc
     1141 cacaacacca cacatatatg gtacacgtgc catatgcgct tgtctcaatt cctgtccgat
     1201 gaagttttca ttgtcaatgt gacggaccag tctggcaaca actcccaaga gtgtggcagc
     1261 tttgtcctgg ctgagagcat caaaccagct ccccccttga acgtgactgt ggccttctca
     1321 ggacgctatg atatctcctg ggactcagct tatgacgaac cctccaacta cgtgctgagg
     1381 ggcaagctac aatatgagct gcagtatcgg aacctcagag acccctatgc tgtgaggccg
     1441 gtgaccaagc tgatctcagt ggactcaaga aacgtctctc ttctccctga agagttccac
     1501 aaagattcta gctaccagct gcaggtgcgg gcagcgcctc agccaggcac ttcattcagg
     1561 gggacctgga gtgagtggag tgaccccgtc atctttcaga cccaggctgg ggagcccgag
     1621 gcaggctggg accctcacat gctgctgctc ctggctgtct tgatcattgt cctggttttc
     1681 atgggtctga agatccacct gccttggagg ctatggaaaa agatatgggc accagtgccc
     1741 acccctgaga gtttcttcca gcccctgtac agggagcaca gcgggaactt caagaaatgg
     1801 gttaataccc ctttcacggc ctccagcata gagttggtgc cacagagttc cacaacaaca
     1861 tcagccttac atctgtcatt gtatccagcc aaggagaaga agttcccggg gctgccgggt
     1921 ctggaagagc aactggagtg tgatggaatg tctgagcctg gtcactggtg cataatcccc
     1981 ttggcagctg gccaagcggt ctcagcctac agtgaggaga gagaccggcc atatggtctg
     2041 gtgtccattg acacagtgac tgtgggagat gcagagggcc tgtgtgtctg gccctgtagc
     2101 tgtgaggatg atggctatcc agccatgaac ctggatgctg gccgagagtc tggccctaat
     2161 tcagaggatc tgctcttggt cacagaccct gcttttctgt cttgcggctg tgtctcaggt
     2221 agtggtctca ggcttggagg ctccccaggc agcctactgg acaggttgag gctgtcattt
     2281 gcaaaggaag gggactggac agcagaccca acctggagaa ctgggtcccc aggagggggc
     2341 tctgagagtg aagcaggttc cccccctggt ctggacatgg acacatttga cagtggcttt
     2401 gcaggttcag actgtggcag ccccgtggag actgatgaag gaccccctcg aagctatctc
     2461 cgccagtggg tggtcaggac ccctccacct gtggacagtg gagcccagag cagcctcgag
     2521 gattacaagg atgacgacga taagggatcc agcgcctggt cccaccctca gtttgagaag
     2581 ggcggaggct ctggcggcgg aagcggagga tctgcttgga gccaccccca gttcgaaaag
     2641 tgatgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2701 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2761 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2821 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2881 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2941 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3001 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3061 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3121 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3181 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3241 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3301 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3361 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3421 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3601 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3661 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3721 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3781 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3841 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3901 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3961 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4021 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4081 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4141 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4201 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4261 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4321 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4381 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4441 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4501 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4561 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4621 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4681 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4741 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4801 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4861 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4981 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5041 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     5881 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5941 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6001 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6061 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7021 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
