LOCUS       V027337                 7328 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027337
VERSION     V027337
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             941..2818
                     /label="SAMHD1(NM_015474)"
                     /note="SAMHD1(NM_015474)"
                     /gene="SAMHD1"
     CDS             2825..2890
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2928..3152
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3198..3626
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3640..3969
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4036..4830
                     /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    5004..5125
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5174..5190)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5198..5214
                     /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(5222..5252)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5267..5288
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5576..6161)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6332..7192)
                     /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(7193..7297)
                     /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 aagcttggta ccgagctcgg atccgccacc atgcagcgag ccgattccga
      961 gcagccctcc aagcgtcccc gttgcgatga cagcccgaga accccctcaa acaccccttc
     1021 cgcagaggca gactggtccc cgggcctgga actccatccc gactacaaga catggggtcc
     1081 ggagcaggtg tgctccttcc tcaggcgcgg tggctttgaa gagccggtgc tgctgaagaa
     1141 catccgagaa aatgaaatca caggcgcatt actgccttgt cttgatgagt ctcgttttga
     1201 aaatcttgga gtaagttcct tgggggagag gaagaagctg cttagttata tccagcgatt
     1261 ggttcaaatc cacgttgata caatgaaggt aattaatgat cctatccatg gccacattga
     1321 gctccaccct ctcctcgtcc gaatcattga tacacctcaa tttcaacgtc ttcgatacat
     1381 caaacagctg ggaggtggtt actatgtttt tccaggagct tcacacaatc gatttgagca
     1441 tagtctaggg gtggggtatc tagcaggatg tctagttcac gcactgggtg aaaaacaacc
     1501 agagctgcag ataagtgaac gagatgttct ctgtgttcag attgctggac tttgtcatga
     1561 tctcggtcat gggccatttt ctcacatgtt tgatggacga tttattccac ttgctcgccc
     1621 ggaggtgaaa tggacgcatg aacaaggctc agttatgatg tttgagcacc ttattaattc
     1681 taatggaatt aagcctgtca tggaacaata tggtctcatc cctgaagaag atatttgctt
     1741 tataaaggaa caaattgtag gaccacttga atcacctgtc gaagattcat tgtggccata
     1801 taaagggcgt cctgaaaaca aaagcttcct ttatgagata gtatctaata aaagaaatgg
     1861 cattgatgtg gacaaatggg attattttgc cagggactgc catcatcttg gaatccaaaa
     1921 taattttgat tacaagcgct ttattaagtt tgcccgtgtc tgtgaagtag acaatgagtt
     1981 gcgtatttgt gctagagata aggaagttgg aaatctgtat gacatgttcc acactcgcaa
     2041 ctctttacac cgtagagctt atcaacacaa agttggcaac attattgata caatgattac
     2101 agatgctttc ctcaaagcag atgactacat agagattaca ggtgctggag gaaaaaagta
     2161 tcgcatttct acagcaattg acgacatgga agcctatact aagctgacag ataacatttt
     2221 tctggagatt ttatactcta ctgatcccaa attgaaagac gcacgagaga ttttaaaaca
     2281 aattgaatac cgtaatctat tcaagtatgt gggtgagacg cagccaacag gacaaataaa
     2341 gattaaaagg gaggactatg aatctcttcc aaaagaggtt gccagtgcta aacccaaagt
     2401 attgctagac gtgaaactga aggctgaaga ttttatagtg gatgttatca acatggatta
     2461 tggaatgcaa gaaaagaatc caattgatca tgttagcttc tattgtaaga ctgcccccaa
     2521 cagagcaatc aggattacta aaaaccaggt ttcacaactt ctgccagaga aatttgcaga
     2581 gcagctgatt cgagtatatt gtaagaaggt ggacagaaag agtttgtatg ccgcaagaca
     2641 atattttgtt cagtggtgtg cagacagaaa tttcaccaag ccgcaggatg gcgatgttat
     2701 agccccactc ataacacctc aaaaaaagga atggaacgac agtacttcag tccaaaatcc
     2761 aactcgcctc cgagaagcat ccaaaagcag agtccagctt tttaaagatg acccaatgct
     2821 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2881 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2941 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3001 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3061 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3121 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3181 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3241 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3301 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3361 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3421 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3481 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3541 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3601 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3841 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3901 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3961 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4021 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4081 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4141 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4201 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4261 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4321 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4381 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4441 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4501 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4561 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4621 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4681 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4741 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4801 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4861 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4921 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4981 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5041 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5101 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5161 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5221 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5281 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5341 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5401 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5461 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5521 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6001 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     6121 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6181 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7321 ctgacgtc
//
