LOCUS       sequence-261744-        5193 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 5193)
  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 5193)
  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: pF141 pAcGFP1 SOD1WT
FEATURES             Location/Qualifiers
     source          1..5193
                     /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
     promoter        2637..2994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2637..2828
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2660..2680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2845..2980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3029..3823
                     /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             3029..3823
                     /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(3083..3102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3693..3712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4011..4030)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4055..4102
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4431..5019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4920..4939
                     /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 agatggtgtg 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 cagcaaccag gtgtggaaag
     2761 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2821 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2881 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2941 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3001 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3061 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3121 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3181 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3241 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3301 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3361 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3421 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3481 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3541 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3601 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3661 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3721 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3781 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3841 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3901 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3961 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4021 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4081 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4141 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4201 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4261 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4321 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4381 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4441 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4501 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4561 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4621 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4681 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4741 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4801 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4861 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4921 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4981 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5041 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5101 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     5161 gtcattagtt catagcccat atatggagtt ccg
//
