LOCUS       Exported                6406 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 6406)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     UUID: 3e8e76fa-5032-4f64-9e0c-fc1dce813e23
            Type: Synthetic
            ConfirmedExperimentally: 0
            CreatedBy: Tomas Gregor
            SequenceClass: UNA
            TransformedInto: Unspecified
FEATURES             Location/Qualifiers
     source          1..6406
                     /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..2637
                     /codon_start=1
                     /locus_tag="SHIP2 V5 His Tag"
                     /label=SHIP2 V5 His Tag
                     /translation="MSKTSDQAYIEFESIEAIVKTASRTKFFIEFYSTCLEEYKKSFEN
                     DAQSSDNINFLKVQWSSRQLPTLKPILADIEYLQDQHLLLTVKSMDGYESYGECVVALK
                     SMIGSTAQQFLTFLSHRGEETGNIRGSMKVRVPTERLGTRERLYEWISIDKDEAGAKSK
                     APSVSRGSQEPRSGSRKPAFTEASCPLSRLFEEPEKPPPTGRPPAPPRAAPREEPLTPR
                     LKPEGAPEPEGVAAPPPKNSFNNPAYYVLEGVPHQLLPPEPPSPARAPVPSATKNKVAI
                     TVPAPQLGHHRHPRVGEGSSSDEESGGTLPPPDFPPPPLPDSAIFLPPSLDPLPGPVVR
                     GRGGAEARGPPPPKAHPRPPLPPGPSPASTFLGEVASGDDRSCSVLQMAKTLSEVDYAP
                     AGPARSALLPGPLELQPPRGLPSDYGRPLSFPPPRIRESIQEDLAEEAPCLQGGRASGL
                     GEAGMSAWLRAIGLERYEEGLVHNGWDDLEFLSDITEEDLEEAGVQDPAHKRLLLDTLQ
                     LSKTRTRPLEGKPIPNPLLGLDSTRTGHHHHHH"
     misc_feature    988..2637
                     /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     1966..1976
                     /locus_tag="XL39"
                     /PCR_conditions="primer sequence:ATTAGGACAAGGCTGGTGGG"
                     /label=XL39
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     primer_bind     2320..2337
                     /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;"
     misc_feature    2347..2544
                     /locus_tag="SAM"
                     /label=SAM
     primer_bind     2543..2585
                     /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    2543..2560
                     /locus_tag="V5 F binding part"
                     /label=V5 F binding part
     misc_feature    2545..2560
                     /locus_tag="linker to Myc"
                     /label=linker to Myc
     primer_bind     2560..2580
                     /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;"
     CDS             2566..2607
                     /codon_start=1
                     /locus_tag="V5 tag"
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2566..2585
                     /locus_tag="primer cast V5 F"
                     /label=primer cast V5 F
     primer_bind     2566..2584
                     /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    2615..2637
                     /locus_tag="primer cast V5 R"
                     /label=primer cast V5 R
     primer_bind     2616..2665
                     /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             2617..2634
                     /codon_start=1
                     /locus_tag="6xHis"
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2622..2665
                     /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    2638..2647
                     /locus_tag="insert NheI"
                     /label=insert NheI
     misc_feature    2638..2643
                     /locus_tag="NheI"
                     /label=NheI
     primer_bind     2663..2679
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2680..2697
                     /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    2690..3312
                     /locus_tag="hGH poly(A) signal"
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2753..2772
                     /locus_tag="XL39(1)"
                     /PCR_conditions="primer sequence:ATTAGGACAAGGCTGGTGGG"
                     /label=XL39
                     /label=XL39(1)
                     /note="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"
     rep_origin      complement(3461..4049)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    4378..4425
                     /locus_tag="HSV TK poly(A) signal"
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal"
     CDS             4657..5451
                     /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        5486..5843
                     /locus_tag="SV40 promoter"
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5500..5635
                     /direction=BOTH
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5845..5949
                     /gene="<i>bla</i>"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     rep_origin      join(5976..6406,1..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 cctgcagggc
     2341 gggcgggcca gcgggctggg cgaggcaggc atgagtgcct ggctgcgggc catcggcttg
     2401 gagcgctatg aggagggcct ggtgcataat ggctgggacg acctggagtt tctcagtgac
     2461 atcaccgagg aggacttgga ggaggctggg gtgcaggacc cggctcacaa gcgcctcctt
     2521 ctggacaccc tgcagctcag caagacgcgt acgcggccgc tcgagggtaa gcctatccct
     2581 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagct
     2641 agcgtttaaa cggccggccg cggtcatagc tgtttcctga acagatcccg ggtggcatcc
     2701 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2761 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2821 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2881 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2941 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3001 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3061 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3121 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata
     3181 actataccag caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt
     3241 gggacatttg agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat
     3301 gcctgttgaa ttgggtacgc ggccagcggc gagcggtatc agctcactca aaggcggtaa
     3361 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3421 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3481 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3541 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3601 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3661 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3721 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3781 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3841 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3901 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3961 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4021 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4081 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4141 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4201 agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt
     4261 cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat
     4321 ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa
     4381 caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc
     4441 cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact
     4501 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     4561 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     4621 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4681 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     4741 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     4801 gcgatccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     4861 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     4921 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     4981 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     5041 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5101 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5161 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     5221 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     5281 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     5341 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5401 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5461 tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc
     5521 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     5581 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     5641 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     5701 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     5761 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     5821 acagctggtt ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca
     5881 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5941 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     6001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6181 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6301 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aatttt
//
