LOCUS       sequence-258461-        5121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5121)
  AUTHORS   Stevens JC, Chia R, Hendriks WT, Bros-Facer V, van Minnen J, Martin 
            JE, Jackson GS, Greensmith L, Schiavo G, Fisher EM
  TITLE     Modification of superoxide dismutase 1 (SOD1) properties by a GFP 
            tag--implications for research into amyotrophic lateral sclerosis 
            (ALS).
  JOURNAL   PLoS One. 2010 . 5(3):e9541.
  PUBMED    20221404
REFERENCE   2  (bases 1 to 5121)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One. 
            2010 . 5(3):e9541."
COMMENT     Sequence Label: pF145 pAcGFP1 SOD1G93A
FEATURES             Location/Qualifiers
     source          1..5121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1269
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1921..2042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1958..1977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2012..2031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2049..2504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2186..2207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2398..2417
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2531..2635
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2660..2680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2704..2914
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2726..2922
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2773..2908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2957..3751
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2957..3751
                     /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(3011..3030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3621..3640
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3939..3958)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3983..4030
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4359..4947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4848..4867
                     /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 gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgcc gagctgttca ccggcatcgt gcccatcctg
      601 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg
      721 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc
      781 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag
      841 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag
      901 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac
      961 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac
     1021 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc
     1081 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg
     1141 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc
     1201 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attccatggc gacgaaggcc
     1321 gtgtgcgtgc tgaagggcga cggcccagtg cagggcatca tcaatttcga gcagaaggaa
     1381 agtaatggac cagtgaaggt gtggggaagc attaaaggac tgactgaagg cctgcatgga
     1441 ttccatgttc atgagtttgg agataataca gcaggctgta ccagtgcagg tcctcacttt
     1501 aatcctctat ccagaaaaca cggtgggcca aaggatgaag agaggcatgt tggagacttg
     1561 ggcaatgtga ctgctgacaa agatgctgtg gccgatgtgt ctattgaaga ttctgtgatc
     1621 tcactctcag gagaccattg catcattggc cgcacactgg tggtccatga aaaagcagat
     1681 gacttgggca aaggtggaaa tgaagaaagt acaaagacag gaaacgctgg aagtcgtttg
     1741 gcttgtggtg taattgggat cgcccaataa gtcgacggta ccgcgggccc gggatccacc
     1801 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1861 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1921 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1981 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2041 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2101 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2161 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2221 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2281 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2341 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2401 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2461 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2521 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2581 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     2641 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2821 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2881 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2941 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3001 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3061 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3121 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3181 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3241 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3301 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3361 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3421 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3481 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3541 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3601 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3661 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3721 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3781 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3841 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3901 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3961 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4021 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4081 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4141 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4201 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4261 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4321 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4381 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4441 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4501 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4561 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4621 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4681 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4741 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4801 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4861 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4921 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4981 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5041 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     5101 tagcccatat atggagttcc g
//
