LOCUS       V028624                 7438 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028624
VERSION     V028624
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1153..2958
                     /label="IGSF10(NM_001178145)"
                     /note="IGSF10(NM_001178145)"
                     /gene="IGSF10"
     polyA_signal    3038..3262
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3308..3736
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3750..4079
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4146..4940
                     /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    5114..5235
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5284..5300)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5308..5324
                     /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(5332..5362)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5377..5398
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5686..6271)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6442..7302)
                     /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(7303..7407)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat ggtggaaagc
     1021 gtaaggcttg agaaccagcc ctgtgaccta agagtgggca gctggatcca cgtctaccct
     1081 aatggatccc tgtttattgg atcagtaaca gaaaaagaca gtggtgtcta cttgtgtgtg
     1141 gcaagaaaca aaatggggga tgatctgata ctgatgcatg ttagcctaag actgaaacct
     1201 gccaaaattg accacaagca gtattttaga aagcaagtgc tccatgggaa agatttccaa
     1261 gtagattgca aagcttccgg ctccccagtg ccagagatat cttggagttt gcctgatgga
     1321 accatgatca acaatgcaat gcaagccgat gacagtggcc acaggactag gagatatacc
     1381 cttttcaaca atggaacttt atacttcaac aaagttgggg tagcggagga aggagattat
     1441 acttgctatg cccagaacac cctagggaaa gatgaaatga aggtccactt aacagttata
     1501 acagctgctc cccggataag gcagagtaac aaaaccaaca agagaatcaa agctggagac
     1561 acagctgtcc ttgactgtga ggtcactggg gatcccaaac caaaaatatt ttggttgctg
     1621 ccttccaatg acatgatttc cttctccatt gataggtaca catttcatgc caatgggtct
     1681 ttgaccatca acaaagtgaa actgctcgat tctggagagt acgtatgtgt agcccgaaat
     1741 cccagtgggg atgacaccaa aatgtacaaa ctggatgtgg tctctaaacc tccattaatc
     1801 aatggtctgt atacaaacag aactgttatt aaagccacag ctgtgagaca ttccaaaaaa
     1861 cactttgact gcagagctga agggacacca tctcctgaag tcatgtggat catgccagac
     1921 aatattttcc tcacagcccc atactatgga agcagaatca cagtccataa aaatggaacc
     1981 ttggaaatta ggaatgtgag gctttcagat tcagccgact ttatctgtgt ggcccgaaat
     2041 gaaggtggag agagcgtgtt ggtagtacag ttagaagtac tggaaatgct gagaagaccg
     2101 acatttagaa atccatttaa tgaaaaaata gttgcccagc tgggaaagtc cacagcattg
     2161 aattgctctg ttgatggtaa cccaccacct gaaataatct ggattttacc aaatggcaca
     2221 cgattttcca atggaccaca aagttatcag tatctgatag caagcaatgg ttcttttatc
     2281 atttctaaaa caactcggga ggatgcagga aaatatcgct gtgcagctag gaataaagtt
     2341 ggctatattg agaaattagt catattagaa attggccaga agccagttat tcttacctat
     2401 gcaccaggga cagtaaaagg catcagtgga gaatctctat cactgcattg tgtgtctgat
     2461 ggaatcccta agccaaatat caaatggact atgccaagtg gttatgtagt agacaggcct
     2521 caaattaatg ggaaatacat attgcatgac aatggcacct tagtcattaa agaagcaaca
     2581 gcttatgaca gaggaaacta tatctgtaag gctcaaaata gtgttggtca tacactgatt
     2641 actgttccag taatgattgt agcctaccct ccccgaatta caaatcgtcc acccaggagt
     2701 attgtcacca ggacaggggc agcctttcag ctccactgtg tggccttggg agttcccaag
     2761 ccagaaatca catgggagat gcctgaccac tcccttctct caacggcaag taaagagagg
     2821 acacatggaa gtgagcagct tcacttacaa ggtaccctag tcattcagaa tccccaaacc
     2881 tccgattctg ggatatacaa atgcacagca aagaacccac ttggtagtga ttatgcagca
     2941 acgtatattc aagtaatctg agaattctgc agatatccag cacagtggcg gccgctcgag
     3001 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3061 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3121 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3181 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3241 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3301 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3361 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3601 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3661 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3721 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3781 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3901 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3961 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4021 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4081 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4141 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4201 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4261 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4321 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4381 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4441 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4501 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4561 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4621 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4681 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4741 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4801 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4861 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4921 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4981 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5041 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5101 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5161 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5221 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5281 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5341 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5401 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5461 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5521 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5581 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5641 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5701 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5761 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5821 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5881 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5941 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6001 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6061 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6121 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6181 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     6241 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6301 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6361 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6421 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7381 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
