LOCUS       V041750                 7945 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041750
VERSION     V041750
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             964..3435
                     /label="DDX20(NM_007204)"
                     /note="DDX20(NM_007204)"
                     /gene="DDX20"
     CDS             3442..3507
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3545..3769
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3815..4243
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4257..4586
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4653..5447
                     /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    5621..5742
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5791..5807)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5815..5831
                     /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(5839..5869)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5884..5905
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6193..6778)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6949..7809)
                     /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(7810..7914)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggcgg cggcatttga agcctcggga gccttagcag cagtggcgac tgctatgccg
     1021 gctgagcatg tggccgtgca ggtcccggcc ccagagccaa cacccgggcc tgtgaggatc
     1081 ctgcggaccg ctcaggatct cagcagcccg cggacccgca cgggggatgt gctgttggcg
     1141 gagccggccg acttcgagtc actgctgctt tcgcggccgg tgctggaggg gctgcgggcg
     1201 gccggcttcg agaggccctc gccggtgcag ctcaaggcca tcccgttggg gcgctgcggg
     1261 ctcgatttaa ttgttcaagc taaatctggc accgggaaaa cctgtgtgtt ctccaccata
     1321 gctttggact ctcttgttct tgaaaactta agtacccaga ttttgatctt ggctcctaca
     1381 agagaaattg ctgtacagat acattctgtt attacagcca ttggaataaa aatggaaggc
     1441 ttagagtgtc atgtctttat tggagggacc ccattatcac aagacaaaac cagacttaaa
     1501 aagtgtcata ttgctgttgg atctcctggc agaattaagc aactcataga acttgactac
     1561 ttgaacccag gcagtatacg cctctttatt cttgatgaag cagataagct tttagaagaa
     1621 ggcagcttcc aggagcaaat aaattggatt tattcttcct tgcctgccag taaacagatg
     1681 ctggcagtat cagctactta tcccgaattt ttggctaatg ctttgacaaa gtacatgaga
     1741 gatcccactt ttgtaagact gaattccagt gatccaagtc tcataggttt gaagcagtat
     1801 tacaaagttg tcaattcata ccctttggca cataaggttt ttgaggaaaa gactcagcat
     1861 ttacaggaac tgttcagcag aattccattt aatcaagctt tagtcttttc taatttgcac
     1921 agcagagcac aacatttggc tgatatcctt tcttctaaag gctttcctgc tgagtgcatt
     1981 tcaggcaata tgaatcagaa tcagcgtctt gatgctatgg ctaaactgaa gcactttcat
     2041 tgcagagtcc tcatttccac agatttgact tctcgtggga ttgatgctga gaaggtgaat
     2101 ctggttgtaa atctggatgt accattggat tgggagacat acatgcatcg gattgggaga
     2161 gctggccgtt ttggtacatt ggggctgaca gtgacctact gttgccgggg agaggaagaa
     2221 aatatgatga tgagaattgc ccagaaatgt aatatcaacc ttctcccttt accagatccc
     2281 attccttctg gtctgatgga agaatgtgtg gattgggatg tggaagttaa agctgctgtg
     2341 catacatatg gtatagcaag tgtacctaac caacccttaa aaaagcaaat tcagaaaata
     2401 gagagaaccc ttcaaattca gaaagctcat ggtgaccaca tggcttcctc tagaaataat
     2461 tctgtatctg gactatcagt caaatcaaaa aataatacca aacaaaagct tcctgtgaaa
     2521 agccactcag aatgtggaat catagaaaaa gcaacgtcac caaaagaact gggctgtgac
     2581 aggcaatccg aagagcaaat gaagaattct gttcagactc ccgttgaaaa ctccaccaac
     2641 agtcagcacc aggtcaaaga agctttacct gtgtcactcc cccagattcc ttgtctgtct
     2701 tcctttaaaa tccatcagcc atacacgttg acttttgctg aattggtaga ggattatgaa
     2761 cattatatta aagaggggtt agagaaacct gtggaaatca tcaggcacta cacaggccct
     2821 ggggatcaga ctgtgaatcc tcaaaatggt tttgtgagaa ataaagttat tgaacagaga
     2881 gtccctgtgt tggcaagtag tagccaatct ggagactctg agagtgacag tgattcttac
     2941 agctcaagaa cctcttccca gagcaaagga aataagtcat acttggaagg ctcttctgat
     3001 aatcagctga aagactctga atctacgcct gtggatgatc gtatttcttt ggaacaacca
     3061 ccaaatggaa gtgacacccc caatccagag aaatatcaag aatcacctgg aatccagatg
     3121 aagacaagac ttaaagaggg ggctagccag agagctaagc agagccggag aaacctaccc
     3181 aggcggtctt ccttcagatt gcagactgaa gcccaggaag atgattggta tgactgtcat
     3241 agggaaatac gtctgagttt ttctgatacc tatcaggatt atgaggagta ctggagagct
     3301 tactacaggg catggcaaga atattatgct gccgcttctc attcatatta ttggaatgct
     3361 cagagacatc caagttggat ggcagcttat cacatgaata ccatttatct acaagaaatg
     3421 atgcatagta accagctcga ggactacaaa gaccatgacg gtgattataa agatcatgac
     3481 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc
     3541 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3601 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3661 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3721 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3781 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3841 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3901 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3961 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4021 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4141 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4261 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4321 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4441 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4501 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4561 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4621 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4681 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4741 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4801 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4861 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4921 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4981 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5041 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5101 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5161 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5221 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5281 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5341 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5401 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5461 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5521 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5581 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5641 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5701 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5761 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5821 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5881 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5941 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6001 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6061 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6121 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6181 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6241 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6301 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6361 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6421 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6481 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6541 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6601 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6661 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6721 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7921 tccccgaaaa gtgccacctg acgtc
//
