LOCUS       Exported                7701 bp ds-DNA     circular SYN 22-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7701)
  AUTHORS   Tomas Gregor
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7701
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     839..856
                     /label=VP1.5
     primer_bind     894..912
                     /label=ColSH F1
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     975..994
                     /label=SHIP2 F
     misc_feature    1027..1032
                     /label=NcoI
     CDS             1029..3932
                     /codon_start=1
                     /label=SHIP2 V5 His Tag
                     /translation="MASACGAPGPGGALGSQAPSWYHRDLSRAAAEELLARAGRDGSFL
                     VRDSESVAGAFALCVLYQKHVHTYRILPDGEDFLAVQTSQGVPVRRFQTLGELIGLYAQ
                     PNQGLVCALLLPVEGEREPDPPDDRDASDGEDEKPPLPPRSGSTSISAPTGPSSPLPAP
                     ETPTAPAAESAPNGLSTVSHDYLKGSYGLDLEAVRGGASHLPHLTRTLATSCRRLHSEV
                     DKVLSGLEILSKVFDQQSSPMVTRLLQQQNLPQTGEQELESLVLKLSVLKDFLSGIQKK
                     ALKALQDMSSTAPPAPQPSTRKAKTIPVQAFEVKLDVTLGDLTKIGKSQKFTLSVDVEG
                     GRLVLLRRQRDSQEDWTTFTHDRIRQLIKSQRVQNKLGVVFEKEKDRTQRKDFIFVSAR
                     KREAFCQLLQLMKNKHSKQSKTSDQAYIEFESIEAIVKTASRTKFFIEFYSTCLEEYKK
                     SFENDAQSSDNINFLKVQWSSRQLPTLKPILADIEYLQDQHLLLTVKSMDGYESYGECV
                     VALKSMIGSTAQQFLTFLSHRGEETGNIRGSMKVRVPTERLGTRERLYEWISIDKDEAG
                     AKSKAPSVSRGSQEPRSGSRKPAFTEASCPLSRLFEEPEKPPPTGRPPAPPRAAPREEP
                     LTPRLKPEGAPEPEGVAAPPPKNSFNNPAYYVLEGVPHQLLPPEPPSPARAPVPSATKN
                     KVAITVPAPQLGHHRHPRVGEGSSSDEESGGTLPPPDFPPPPLPDSAIFLPPSLDPLPG
                     PVVRGRGGAEARGPPPPKAHPRPPLPPGPSPASTFLGEVASGDDRSCSVLQMAKTLSEV
                     DYAPAGPARSALLPGPLELQPPRGLPSDYGRPLSFPPPRIRESIQEDLAEEAPCLQGGR
                     ASGLGEAGMSAWLRAIGLERYEEGLVHNGWDDLEFLSDITEEDLEEAGVQDPAHKRLLL
                     DTLQLSKTRTRPLEGKPIPNPLLGLDSTRTGHHHHHH"
     misc_feature    1029..2282
                     /label=1. N-term
     misc_feature    1089..1379
                     /label=SH2 PDB
     misc_feature    1395..2282
                     /label=2. N-term no SH2
     primer_bind     1395..1412
                     /label=PR1 F (SalI)
     primer_bind     1717..1735
                     /label=SeqSH 1
     primer_bind     2148..2166
                     /label=SeqSH 2
     primer_bind     complement(2261..2302)
                     /label=Delta-PHOS R2
     primer_bind     complement(2261..2282)
                     /label=Phos R2
     primer_bind     2263..2304
                     /label=Delta-PHOS F2
     misc_feature    2283..3932
                     /label=5. C-term
     misc_feature    2283..3632
                     /label=6. pro-rich
     primer_bind     2283..2304
                     /label=Phos F2 (SalI)
     primer_bind     2617..2635
                     /label=SeqSH 3
     misc_feature    2868..3374
                     /label=Proline rich
     primer_bind     complement(3615..3632)
                     /label=SAM R
     misc_feature    3642..3839
                     /label=SAM
     primer_bind     3838..3880
                     /label=V5 F
     misc_feature    3838..3855
                     /label=V5 F binding part
     misc_feature    3840..3855
                     /label=linker to Myc
     primer_bind     complement(3855..3875)
                     /label=SHIP2 R
     CDS             3861..3902
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3861..3880
                     /label=primer cast V5 F
     primer_bind     3861..3879
                     /label=V5 Short F
     misc_feature    3910..3932
                     /label=primer cast V5 R
     primer_bind     complement(3911..3960)
                     /label=V5 R
     CDS             3912..3929
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3933..3942
                     /label=insert NheI
     misc_feature    3933..3938
                     /label=NheI
     primer_bind     complement(3958..3974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3975..3992)
                     /label=Seq SHIP2 R
     polyA_signal    3985..4607
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4048..4067)
                     /label=XL39
     rep_origin      complement(4756..5344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    5673..5720
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal"
     CDS             complement(5952..6746)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(6781..7138)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6795..6930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7140..7244)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7271..7701,1..25)
                     /direction=RIGHT
                     /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 attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggcctcggcc tgcggggcgc cgggcccggg gggcgccctg ggcagccagg
     1081 ccccctcctg gtaccaccgc gacctgagcc gggcggccgc ggaggagctg ctggcccggg
     1141 cgggccgcga tggcagcttc ctggtccgag acagcgagag cgtggcgggg gccttcgcgc
     1201 tctgcgtcct gtatcagaag catgtgcaca cgtatcgcat tctgcctgat ggagaagatt
     1261 tcttggctgt gcagacctcg cagggtgtgc ctgtgcgccg cttccagacc ctgggtgagc
     1321 tcatcggcct gtacgcccag cccaaccagg gccttgtgtg cgccctgctt cttcctgtag
     1381 agggtgagcg agagccggac ccaccggatg accgggatgc ctcagatggg gaggatgaga
     1441 agcccccgct gcccccgcgc tctggctcca ccagcatttc tgcccccact gggcccagca
     1501 gtcccctgcc agctcctgag actcccacag ctccagctgc tgagagtgct cccaatgggc
     1561 tgagcaccgt ctcgcacgac tacctgaaag gcagctatgg gctggacctg gaagctgtga
     1621 ggggtggagc cagccacctg ccccacctca cccgtaccct cgctacctca tgccggaggc
     1681 tgcacagtga ggtggacaag gtcctgtcag gcctggagat cctgtccaag gtgtttgacc
     1741 agcagagctc gcccatggtg acccgccttt tgcagcagca gaacctgcca cagacagggg
     1801 agcaggaact agagagcctg gtgctgaagc tgtcagtgct aaaggacttc ctgtcaggca
     1861 tccagaagaa ggccctgaag gccctacagg acatgagctc cacagcaccc ccagctccgc
     1921 agccatccac acgtaaggcc aagaccatcc ccgtgcaggc ctttgaggtg aagctagatg
     1981 tgaccctggg tgacctgacc aagattggga agtcacagaa gttcacgctg agcgtggatg
     2041 tggagggtgg gcggctggtg ctgctgcgga gacagcggga ctcccaggag gactggacca
     2101 ccttcacgca cgaccgcatc cgccagctca ttaagtccca gcgtgtccag aacaagctgg
     2161 gtgttgtgtt tgagaaggag aaggaccgga ctcagcgcaa ggacttcatc tttgtcagtg
     2221 cccggaagcg ggaggccttc tgccagctgt tgcagctcat gaagaacaag cactccaagc
     2281 agtcaaagac ttcagaccag gcctacattg agtttgagag catcgaggcc attgtgaaga
     2341 cagccagccg caccaagttc ttcatcgagt tctactctac ctgcctggag gaatacaaga
     2401 agagctttga gaatgatgcc cagagcagtg acaacatcaa cttcctcaaa gtgcagtggt
     2461 cttcacgcca gctgcccacg ctcaaaccaa ttctggctga tatcgagtac ctgcaggacc
     2521 agcacctcct gctcacagtc aagtccatgg atggctatga atcctatggg gagtgtgtgg
     2581 ttgcactcaa atccatgatc ggcagcacgg cccaacagtt cctgaccttc ctatcccacc
     2641 gtggcgagga gacaggcaat atcagaggct ccatgaaggt gcgggtgccc acggagcgcc
     2701 tgggcacccg tgagcggctc tacgagtgga tcagcattga taaggatgag gcaggagcaa
     2761 agagcaaagc cccctctgtg tcccgaggga gccaggagcc caggtcaggg agccgcaagc
     2821 cagccttcac agaggcctcc tgcccgctct ccaggttatt tgaagaacca gagaaaccgc
     2881 caccaacggg gaggccccca gccccacccc gagcagctcc ccgggaggag cccttgaccc
     2941 ccaggttgaa gccagaggga gctcctgaac cagaaggggt ggcggccccc ccacccaaga
     3001 acagcttcaa taaccctgcc tactacgtcc ttgaaggggt cccgcaccag ctgctgcccc
     3061 cggagccacc ctcgcctgcc agggcccctg tcccatctgc caccaagaac aaagtggcca
     3121 ttacagtgcc tgctccacag cttgggcacc accggcaccc tcgtgtggga gaggggagtt
     3181 cttcagatga ggagtctgga ggcacactgc cccctccaga ctttccacct ccaccactgc
     3241 cggactcagc catcttcctg ccccccagcc tggatccttt accagggcca gtggtccggg
     3301 gccgtggtgg ggctgaggcc cgtggcccac cacctcccaa ggcccatcca aggcctccac
     3361 tgcccccagg cccctcacca gccagcactt tcctggggga agtggccagt ggggatgacc
     3421 ggtcctgctc ggtgctgcag atggccaaga cgctgagcga ggtggactat gcccctgctg
     3481 ggcctgcacg ctcagcgctc ctcccaggcc ccctggagct gcagcccccc cggggactgc
     3541 cctcggacta tggccggccc ctcagcttcc ctccaccccg catccgggag agcatccagg
     3601 aagacctggc agaggaggct ccgtgcctgc agggcgggcg ggccagcggg ctgggcgagg
     3661 caggcatgag tgcctggctg cgggccatcg gcttggagcg ctatgaggag ggcctggtgc
     3721 ataatggctg ggacgacctg gagtttctca gtgacatcac cgaggaggac ttggaggagg
     3781 ctggggtgca ggacccggct cacaagcgcc tccttctgga caccctgcag ctcagcaaga
     3841 cgcgtacgcg gccgctcgag ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     3901 cgcgtaccgg tcatcatcac catcaccatt gagctagcgt ttaaacggcc ggccgcggtc
     3961 atagctgttt cctgaacaga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     4021 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     4081 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     4141 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     4201 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     4261 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     4321 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     4381 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4441 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     4501 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     4561 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     4621 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4681 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4741 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4801 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4861 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4921 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4981 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5041 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5101 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5161 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5221 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5281 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5341 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5401 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5461 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     5521 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     5581 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     5641 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     5701 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     5761 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     5821 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     5881 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     5941 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     6001 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     6061 gcaatatcac gggtagccaa cgctatgtcc tgatagcgat ccgccacacc cagccggcca
     6121 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     6181 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     6241 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     6301 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     6361 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     6421 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     6481 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     6541 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     6601 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     6661 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     6721 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     6781 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     6841 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     6901 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     6961 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     7021 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     7081 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     7141 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7201 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7261 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7321 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7381 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7441 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7501 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7561 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7621 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7681 caaaaattta acgcgaattt t
//
