LOCUS       V035928                 6431 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035928
VERSION     V035928
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..1924
                     /label="ERLIN2(NM_007175)"
                     /note="ERLIN2(NM_007175)"
                     /gene="ERLIN2"
     CDS             1934..1951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    1998..2222
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2268..2696
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2710..3039
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3106..3900
                     /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    4074..4195
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4244..4260)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4268..4284
                     /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(4292..4322)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4337..4358
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4646..5231)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5402..6262)
                     /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(6263..6367)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gctcagttgg gagcagttgt ggctgtggct tccagtttct tttgtgcatc
      961 tctcttctca gctgtgcaca agatagaaga gggacatatt ggggtatatt acagaggcgg
     1021 tgccctgctg acttcgacca gcggccctgg tttccatctc atgctccctt tcatcacatc
     1081 atataagtct gtgcagacca cactccagac agatgaggtg aagaatgtac cttgtgggac
     1141 tagtggtggt gtgatgatct actttgacag aattgaagtg gtgaacttcc tggtcccgaa
     1201 cgcagtgtat gatatagtga agaactatac tgctgactat gacaaggccc tcatcttcaa
     1261 caagatccac cacgaactga accagttctg cagtgtgcac acgcttcaag aggtctacat
     1321 tgagctgttt gatcagattg atgaaaatct caaactggct ttgcaacagg acctgacctc
     1381 catggcccct gggctggtca ttcaagctgt gcgggtaaca aagcccaaca taccagaggc
     1441 aatccgcaga aactacgagt tgatggaaag tgagaagaca aagcttctca ttgccgccca
     1501 gaaacagaag gtggtggaaa aggaagcaga gacagagcgg aagaaggcgc tcattgaggc
     1561 agaaaaagtg gcccaggtgg ctgagatcac ctacgggcag aaggtgatgg agaaggagac
     1621 tgagaagaag atttcagaaa ttgaagatgc tgcatttctg gcccgggaga aggcaaaggc
     1681 agatgctgag tgctacactg ctatgaaaat agccgaagcc aataagctga agctaacccc
     1741 tgaatatctg cagctgatga agtacaaggc cattgcttcc aacagcaaga tttactttgg
     1801 caaagacatt cctaacatgt tcatggactc tgcgggcagt gtgagcaagc agtttgaggg
     1861 gctagctgac aagctaagct ttggcttaga agatgaaccc ttggagacgg ccactaagga
     1921 gaatgcggcc gcacatcacc atcatcacca ttgactcgag tctagagggc ccgtttaaac
     1981 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2161 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2221 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2281 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2341 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2401 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2461 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2521 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2581 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2641 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2941 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3061 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3121 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3181 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3241 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3301 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3361 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3421 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3481 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3541 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3601 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3661 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3721 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3781 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3841 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3901 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3961 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4021 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4081 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4141 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4201 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4261 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4321 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4381 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4441 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4501 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4561 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4621 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4681 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4741 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4801 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4861 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4921 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4981 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5041 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5101 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5161 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5701 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6001 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6361 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6421 tcccctatgg t
//
