LOCUS       Exported                5722 bp ds-DNA     circular SYN 11-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5722)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Type: Synthetic
            ConfirmedExperimentally: 0
            Created: 2016.6.14
            LastModified: 2016.9.22
            CreatedBy: Tomas Gregor
            TransformedInto: Unspecified
FEATURES             Location/Qualifiers
     source          1..5722
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     166..182
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /locus_tag="CMV enhancer"
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /locus_tag="CMV promoter"
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     839..856
                     /locus_tag="VP1.5"
                     /PCR_conditions="primer sequence:GGACTTTCCAAAATGTCG"
                     /label=VP1.5
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     894..912
                     /locus_tag="ColSH F1"
                     /PCR_conditions="primer sequence:CGTGTACGGTGGGAGGTCT"
                     /label=ColSH F1
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     promoter        952..970
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     979..1005
                     /locus_tag="PR1 F EcoRI"
                     /PCR_conditions="primer
                     sequence:TAGACTGAATTCCCGGACCCACCGGATGAC"
                     /label=PR1 F EcoRI
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     CDS             985..1953
                     /codon_start=1
                     /locus_tag="SHIP2 V5 His Tag"
                     /label=SHIP2 V5 His Tag
                     /translation="MPDPPDDRDASDGEDEKPPLPPRSGSTSISAPTGPSSPLPAPETP
                     TAPAAESAPNGLSTVSHDYLKGSYGLDLEAVRGGASHLPHLTRTLATSCRRLHSEVDKV
                     LSGLEILSKVFDQQSSPMVTRLLQQQNLPQTGEQELESLVLKLSVLKDFLSGIQKKALK
                     ALQDMSSTAPPAPQPSTRKAKTIPVQAFEVKLDVTLGDLTKIGKSQKFTLSVDVEGGRL
                     VLLRRQRDSQEDWTTFTHDRIRQLIKSQRVQNKLGVVFEKEKDRTQRKDFIFVSARKRE
                     AFCQLLQLMKNKHSKQLEGKPIPNPLLGLDSTRTGHHHHHH"
     misc_feature    988..1875
                     /locus_tag="1. N-term"
                     /label=1. N-term
     misc_feature    988..1875
                     /locus_tag="2. N-term no SH2"
                     /label=2. N-term no SH2
     primer_bind     1310..1328
                     /locus_tag="SeqSH 1"
                     /PCR_conditions="primer sequence:AGATCCTGTCCAAGGTGTT"
                     /label=SeqSH 1
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     1741..1759
                     /locus_tag="SeqSH 2"
                     /PCR_conditions="primer sequence:CAGAACAAGCTGGGTGTTG"
                     /label=SeqSH 2
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     1854..1881
                     /locus_tag="Phos R2"
                     /PCR_conditions="primer
                     sequence:TCAATTCTCGAGCTGCTTGGAGTGCTTGTTCTTC"
                     /label=Phos R2
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     1854..1875
                     /locus_tag="IPHOS R2"
                     /PCR_conditions="primer 
                     sequence:GCCTGGTCTGAAGTCTTTGACTGCTTGGAGTGCTTGTTCTTC"
                     /label=IPHOS R2
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     1872..1901
                     /locus_tag="V5 F"
                     /PCR_conditions="primer 
                     sequence:AGACGCGTACGCGGCCGCTCGAGGGTAAGCCTATCCCTAACCC"
                     /label=V5 F
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     misc_feature    1876..1953
                     /locus_tag="5. C-term"
                     /label=5. C-term
     primer_bind     1876..1896
                     /locus_tag="SHIP2 R"
                     /PCR_conditions="primer sequence:AGGGATAGGCTTACCCTCGAG"
                     /label=SHIP2 R
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     misc_feature    1876
                     /locus_tag="V5 F binding part"
                     /label=V5 F binding part
     misc_feature    1876
                     /locus_tag="linker to Myc"
                     /label=linker to Myc
     CDS             1882..1923
                     /codon_start=1
                     /locus_tag="V5 tag"
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1882..1901
                     /locus_tag="primer cast V5 F"
                     /label=primer cast V5 F
     primer_bind     1882..1900
                     /locus_tag="V5 Short F"
                     /PCR_conditions="primer sequence:GGTAAGCCTATCCCTAACC"
                     /label=V5 Short F
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     misc_feature    1931..1953
                     /locus_tag="primer cast V5 R"
                     /label=primer cast V5 R
     primer_bind     1932..1981
                     /locus_tag="V5 R"
                     /PCR_conditions="primer 
                     sequence:GACCGCGGCCGGCCGTTTAAACGCTAGCTCAATGGTGATGGTGATGATG 
                     A"
                     /label=V5 R
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     CDS             1933..1950
                     /codon_start=1
                     /locus_tag="6xHis"
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1938..1981
                     /locus_tag="V5 R(1)"
                     /PCR_conditions="primer 
                     sequence:GACCGCGGCCGGCCGTTTAAACGCTAGCTCAATGGTGATGGTGATGATG 
                     A"
                     /label=V5 R
                     /label=V5 R(1)
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     misc_feature    1954..1963
                     /locus_tag="insert NheI"
                     /label=insert NheI
     misc_feature    1954..1959
                     /locus_tag="NheI"
                     /label=NheI
     primer_bind     1979..1995
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1996..2013
                     /locus_tag="Seq SHIP2 R"
                     /PCR_conditions="primer sequence:TGCCACCCGGGATCTGTT"
                     /label=Seq SHIP2 R
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     polyA_signal    2006..2628
                     /locus_tag="hGH poly(A) signal"
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2069..2088
                     /locus_tag="XL39"
                     /PCR_conditions="primer sequence:ATTAGGACAAGGCTGGTGGG"
                     /label=XL39
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     rep_origin      complement(2777..3365)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    3694..3741
                     /locus_tag="HSV TK poly(A) signal"
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal"
     CDS             3973..4767
                     /codon_start=1
                     /gene="<i>aph(3')-II</i> (or <i>nptII</i>)"
                     /locus_tag="NeoR/KanR"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (GeneticinA(R))"
                     /translation="MEELVKKAIEGDALRIGSGDTVKHEEAVSPFAAKLFSNITGSQRY
                     VLIAIRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILAVGHARLEPGEQFGWREPLM
                     LFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQPPHCISHDGYFLGRSKVR*Q
                     EILPRHFAQ*QPVPSRFSDNVEHSCARNARRGQPR*PRCLVLQFIQGTGQVGLDKKNRA
                     PLR*QPEHGGIRAADCLLCPVIAE*PLHPSGRRTCVQSILFNH"
     promoter        4802..5159
                     /locus_tag="SV40 promoter"
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4816..4951
                     /direction=BOTH
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5161..5265
                     /gene="<i>bla</i>"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     rep_origin      join(5292..5722,1..25)
                     /direction=RIGHT
                     /locus_tag="f1 ori"
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcatgccg gacccaccgg atgaccggga tgcctcagat
     1021 ggggaggatg agaagccccc gctgcccccg cgctctggct ccaccagcat ttctgccccc
     1081 actgggccca gcagtcccct gccagctcct gagactccca cagctccagc tgctgagagt
     1141 gctcccaatg ggctgagcac cgtctcgcac gactacctga aaggcagcta tgggctggac
     1201 ctggaagctg tgaggggtgg agccagccac ctgccccacc tcacccgtac cctcgctacc
     1261 tcatgccgga ggctgcacag tgaggtggac aaggtcctgt caggcctgga gatcctgtcc
     1321 aaggtgtttg accagcagag ctcgcccatg gtgacccgcc ttttgcagca gcagaacctg
     1381 ccacagacag gggagcagga actagagagc ctggtgctga agctgtcagt gctaaaggac
     1441 ttcctgtcag gcatccagaa gaaggccctg aaggccctac aggacatgag ctccacagca
     1501 cccccagctc cgcagccatc cacacgtaag gccaagacca tccccgtgca ggcctttgag
     1561 gtgaagctag atgtgaccct gggtgacctg accaagattg ggaagtcaca gaagttcacg
     1621 ctgagcgtgg atgtggaggg tgggcggctg gtgctgctgc ggagacagcg ggactcccag
     1681 gaggactgga ccaccttcac gcacgaccgc atccgccagc tcattaagtc ccagcgtgtc
     1741 cagaacaagc tgggtgttgt gtttgagaag gagaaggacc ggactcagcg caaggacttc
     1801 atctttgtca gtgcccggaa gcgggaggcc ttctgccagc tgttgcagct catgaagaac
     1861 aagcactcca agcagctcga gggtaagcct atccctaacc ctctcctcgg tctcgattct
     1921 acgcgtaccg gtcatcatca ccatcaccat tgagctagcg tttaaacggc cggccgcggt
     1981 catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct
     2041 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     2101 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     2161 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     2221 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     2281 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     2341 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     2401 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     2461 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     2521 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     2581 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     2641 agcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2701 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2761 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2821 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2881 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2941 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3001 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3061 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3121 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3181 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3241 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3301 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3361 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3421 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3481 aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc
     3541 ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg
     3601 gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag
     3661 agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt
     3721 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     3781 tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga
     3841 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     3901 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     3961 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4021 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4081 agcaatatca cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc
     4141 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4201 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     4261 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     4321 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     4381 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     4441 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     4501 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     4561 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     4621 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     4681 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     4741 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat
     4801 ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     4861 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     4921 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     4981 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5041 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     5101 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg
     5161 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5221 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5281 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5341 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5401 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5461 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5521 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5581 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5641 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5701 acaaaaattt aacgcgaatt tt
//
