LOCUS       Exported                5863 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses myc-M87L195V spastin isoform in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Tag 3B/myc-M87L195V
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Solowska JM, Garbern JY, Baas PW
  TITLE     Evaluation of loss of function as an explanation for SPG4-based 
            hereditary spastic paraplegia.
  JOURNAL   Hum Mol Genet. 2010 Jul 15;19(14):2767-79. doi: 10.1093/hmg/ddq177. 
            Epub 2010 Apr 29.
  PUBMED    20430936
REFERENCE   2  (bases 1 to 5863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5863
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      604..613
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2298..2317)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2299..2317)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2591..2712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2628..2647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2682..2701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2719..3174)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2856..2877)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3068..3087
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3201..3305
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3307..3664
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3330..3350)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3515..3650
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3577..3596
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3699..4493
                     /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"
     primer_bind     complement(3753..3772)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4363..4382
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4681..4700)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4725..4772
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5101..5689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5590..5609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg
      601 gcggccgcca tggagcagaa actcatctct gaagaggatc tgagcccggg cggatccgcc
      661 gccaagagga gctccggggc cgcgccagca cctgcctcgg cctcggcccc ggcgccggtg
      721 ccgggcggcg aggccgagcg cgtccgagtc ttccacaaac aggccttcga gtacatctcc
      781 attgccctgc gcatcgatga ggatgagaaa gcaggacaga aggagcaagc tgtggaatgg
      841 tataagaaag gtattgaaga actggaaaaa ggaatagctg ttatagttac aggacaaggt
      901 gaacagtgtg aaagagctag acgccttcaa gctaaaatga tgactaattt ggttatggcc
      961 aaggaccgct tacaacttgt agagaagatg caaccagttt tgccattttc caagtcacaa
     1021 acggacgtct ataatgacag tactaacttg gcatgccgca atggacatct ccagtcagaa
     1081 agtggagctg ttccaaaaag aaaagacccc ttaacacaca ctagtaattc actgcctcgt
     1141 tcaaaaacag ttatgaaaac tggatctgca ggcctttcag gccaccatag agcacctagt
     1201 tacagtggtt tatccatggt ttctggagtg aaacagggat ctggtcctgc tcctaccact
     1261 cataagggta ctccgaaaac aaataggaca aataaacctt ctacccctac aactgctact
     1321 cgtaagaaaa aagacttgaa gaattttagg aatgtggaca gcaaccttgc taaccttata
     1381 atgaatgaaa ttgtggacaa tggaacagct gttaaatttg atgatatagc tggtcaagac
     1441 ttggcaaaac aagcattgca agaaattgtt attcttcctt ctctgaggcc tgagttgttc
     1501 acagggctta gagctcctgc cagagggctg ttactctttg gtccacctgg gaatgggaag
     1561 acaatgctgg ctaaagcagt agctgcagaa tcgaatgcaa ccttctttaa tataagtgct
     1621 gcaagtttaa cttcaaaata cgtgggagaa ggagagaaat tggtgagggc tctttttgct
     1681 gtggctcgag aacttcaacc ttctataatt tttatagatg aagttgatag ccttttgtgt
     1741 gaaagaagag aaggggagca cgatgctagt agacgcctaa aaactgaatt tctaatagaa
     1801 tttgatggtg tacagtctgc tggagatgac agagtacttg taatgggtgc aactaatagg
     1861 ccacaagagc ttgatgaggc tgttctcagg cgtttcatca aacgggtata tgtgtcttta
     1921 ccaaatgagg agacaagact acttttgctt aaaaatctgt tatgtaaaca aggaagtcca
     1981 ttgacccaaa aagaactagc acaacttgct agaatgactg atggatactc aggaagtgac
     2041 ctaacagctt tggcaaaaga tgcagcactg ggtcctatcc gagaactaaa accagaacag
     2101 gtgaagaata tgtctgccag tgagatgaga aatattcgat tatctgactt cactgaatcc
     2161 ttgaaaaaaa taaaacgcag cgtcagccct caaactttag aagcgtacat acgttggaac
     2221 aaggactttg gagataccac tgtttaaggg cccggtacct taattaatta aggtaccagg
     2281 taagtgtacc caattcgccc tatagtgagt cgtattacaa ttcactcgat cgcccttccc
     2341 aacagttgcg cagcctgaat ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa
     2401 actactgatt ctaattgttt gtgtatttta gattcacagt cccaaggctc atttcaggcc
     2461 cctcagtcct cacagtctgt tcatgatcat aatcagccat accacatttg tagaggtttt
     2521 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2581 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2641 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2701 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2761 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2821 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2881 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2941 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3001 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3061 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3121 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3181 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3241 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3301 agaatcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3361 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3541 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3601 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3661 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3721 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3781 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3841 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3901 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3961 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4021 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4081 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4141 tggaagccgg tcttgtcgat caggatgatc tggacgaaga acatcagggg ctcgcgccag
     4201 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4261 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4321 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4381 ttgctgaaga acttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4441 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4501 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4561 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4621 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4681 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4741 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4801 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4861 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4921 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4981 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5761 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     5821 caattacggg gtcattagtt catagcccat atatggagtt ccg
//
