LOCUS       Exported                6184 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 6184)
  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..6184
                     /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..1007
                     /locus_tag="Phos F2 EcoRI"
                     /PCR_conditions="primer
                     sequence:GCAGGTGAATTCTCAAAGACTTCAGACCAGGC"
                     /label=Phos F2 EcoRI
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     CDS             985..2415
                     /codon_start=1
                     /locus_tag="SHIP2 V5 His Tag"
                     /label=SHIP2 V5 His Tag
                     /translation="MSKTSDQAYIEFESIEAIVKTASRTKFFIEFYSTCLEEYKKSFEN
                     DAQSSDNINFLKVQWSSRQLPTLKPILADIEYLQDQHLLLTVKSMDGYESYGECVVALK
                     SMIGSTAQQFLTFLSHRGEETGNIRGSMKVRVPTERLGTRERLYEWISIDKDEAGAKSK
                     APSVSRGSQEPRSGSRKPAFTEASCPLSRLFEEPEKPPPTGRPPAPPRAAPREEPLTPR
                     LKPEGAPEPEGVAAPPPKNSFNNPAYYVLEGVPHQLLPPEPPSPARAPVPSATKNKVAI
                     TVPAPQLGHHRHPRVGEGSSSDEESGGTLPPPDFPPPPLPDSAIFLPPSLDPLPGPVVR
                     GRGGAEARGPPPPKAHPRPPLPPGPSPASTFLGEVASGDDRSCSVLQMAKTLSEVDYAP
                     AGPARSALLPGPLELQPPRGLPSDYGRPLSFPPPRIRESIQEDLAEEAPCLQLEGKPIP
                     NPLLGLDSTRTGHHHHHH"
     misc_feature    988..2415
                     /locus_tag="5. C-term"
                     /label=5. C-term
     misc_feature    988..2337
                     /locus_tag="6. pro-rich"
                     /label=6. pro-rich
     primer_bind     988..1009
                     /locus_tag="IPHOS F2"
                     /PCR_conditions="primer 
                     sequence:AGAACAAGCACTCCAAGCAGTCAAAGACTTCAGACCAGGCCT"
                     /label=IPHOS F2
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     1322..1340
                     /locus_tag="SeqSH 3"
                     /PCR_conditions="primer sequence:AGTTCCTGACCTTCCTATC"
                     /label=SeqSH 3
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     misc_feature    1573..2079
                     /locus_tag="Proline rich"
                     /label=Proline rich
     primer_bind     2320..2343
                     /locus_tag="SAM R"
                     /PCR_conditions="primer
                     sequence:TAATATCTCGAGCTGCAGGCACGGAGCCTC"
                     /label=SAM R
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     2334..2363
                     /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;"
     primer_bind     2338..2358
                     /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    2338
                     /locus_tag="linker to Myc"
                     /label=linker to Myc
     misc_feature    2338
                     /locus_tag="V5 F binding part"
                     /label=V5 F binding part
     CDS             2344..2385
                     /codon_start=1
                     /locus_tag="V5 tag"
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2344..2363
                     /locus_tag="primer cast V5 F"
                     /label=primer cast V5 F
     primer_bind     2344..2362
                     /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    2393..2415
                     /locus_tag="primer cast V5 R"
                     /label=primer cast V5 R
     primer_bind     2394..2443
                     /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             2395..2412
                     /codon_start=1
                     /locus_tag="6xHis"
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2400..2443
                     /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    2416..2425
                     /locus_tag="insert NheI"
                     /label=insert NheI
     misc_feature    2416..2421
                     /locus_tag="NheI"
                     /label=NheI
     primer_bind     2441..2457
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2458..2475
                     /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    2468..3090
                     /locus_tag="hGH poly(A) signal"
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2531..2550
                     /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(3239..3827)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    4156..4203
                     /locus_tag="HSV TK poly(A) signal"
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal"
     CDS             4435..5229
                     /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        5264..5621
                     /locus_tag="SV40 promoter"
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5278..5413
                     /direction=BOTH
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5623..5727
                     /gene="<i>bla</i>"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     rep_origin      join(5754..6184,1..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 attcatgtca aagacttcag accaggccta cattgagttt
     1021 gagagcatcg aggccattgt gaagacagcc agccgcacca agttcttcat cgagttctac
     1081 tctacctgcc tggaggaata caagaagagc tttgagaatg atgcccagag cagtgacaac
     1141 atcaacttcc tcaaagtgca gtggtcttca cgccagctgc ccacgctcaa accaattctg
     1201 gctgatatcg agtacctgca ggaccagcac ctcctgctca cagtcaagtc catggatggc
     1261 tatgaatcct atggggagtg tgtggttgca ctcaaatcca tgatcggcag cacggcccaa
     1321 cagttcctga ccttcctatc ccaccgtggc gaggagacag gcaatatcag aggctccatg
     1381 aaggtgcggg tgcccacgga gcgcctgggc acccgtgagc ggctctacga gtggatcagc
     1441 attgataagg atgaggcagg agcaaagagc aaagccccct ctgtgtcccg agggagccag
     1501 gagcccaggt cagggagccg caagccagcc ttcacagagg cctcctgccc gctctccagg
     1561 ttatttgaag aaccagagaa accgccacca acggggaggc ccccagcccc accccgagca
     1621 gctccccggg aggagccctt gacccccagg ttgaagccag agggagctcc tgaaccagaa
     1681 ggggtggcgg cccccccacc caagaacagc ttcaataacc ctgcctacta cgtccttgaa
     1741 ggggtcccgc accagctgct gcccccggag ccaccctcgc ctgccagggc ccctgtccca
     1801 tctgccacca agaacaaagt ggccattaca gtgcctgctc cacagcttgg gcaccaccgg
     1861 caccctcgtg tgggagaggg gagttcttca gatgaggagt ctggaggcac actgccccct
     1921 ccagactttc cacctccacc actgccggac tcagccatct tcctgccccc cagcctggat
     1981 cctttaccag ggccagtggt ccggggccgt ggtggggctg aggcccgtgg cccaccacct
     2041 cccaaggccc atccaaggcc tccactgccc ccaggcccct caccagccag cactttcctg
     2101 ggggaagtgg ccagtgggga tgaccggtcc tgctcggtgc tgcagatggc caagacgctg
     2161 agcgaggtgg actatgcccc tgctgggcct gcacgctcag cgctcctccc aggccccctg
     2221 gagctgcagc ccccccgggg actgccctcg gactatggcc ggcccctcag cttccctcca
     2281 ccccgcatcc gggagagcat ccaggaagac ctggcagagg aggctccgtg cctgcagctc
     2341 gagggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat
     2401 caccatcacc attgagctag cgtttaaacg gccggccgcg gtcatagctg tttcctgaac
     2461 agatcccggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     2521 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     2581 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     2641 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     2701 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     2761 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     2821 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     2881 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     2941 ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc
     3001 tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt
     3061 tgggtccact cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga gcggtatcag
     3121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3901 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3961 caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg
     4021 ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc
     4081 gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     4141 gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg
     4201 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     4261 tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     4321 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     4381 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     4441 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     4501 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     4561 caacgctatg tcctgatagc gatccgccac acccagccgg ccacagtcga tgaatccaga
     4621 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     4681 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     4741 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     4801 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     4861 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     4921 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     4981 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     5041 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     5101 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     5161 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     5221 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc
     5281 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     5341 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     5401 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     5461 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     5521 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5581 ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata
     5641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     5761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5821 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5881 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5941 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6061 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6121 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6181 tttt
//
