LOCUS       sequence-205961-        5187 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 5187)
  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 5187)
  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: pF146 pSOD1WTAcGFP1
FEATURES             Location/Qualifiers
     source          1..5187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        774..1077
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1078..1281
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1232..1252
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1847..2566
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2688..2809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2725..2744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2779..2798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2816..3271)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2953..2974)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3165..3184
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3298..3402
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3404..3761
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3404..3595
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3427..3447)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3612..3747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3796..4590
                     /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             3796..4590
                     /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(3850..3869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4460..4479
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4778..4797)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4822..4869
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
      721 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      781 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      841 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      901 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      961 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     1021 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     1081 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     1141 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     1201 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     1261 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
     1321 cagatctcga gctcaagctt cgaattccat ggcgacgaag gccgtgtgcg tgctgaaggg
     1381 cgacggccca gtgcagggca tcatcaattt cgagcagaag gaaagtaatg gaccagtgaa
     1441 ggtgtgggga agcattaaag gactgactga aggcctgcat ggattccatg ttcatgagtt
     1501 tggagataat acagcaggct gtaccagtgc aggtcctcac tttaatcctc tatccagaaa
     1561 acacggtggg ccaaaggatg aagagaggca tgttggagac ttgggcaatg tgactgctga
     1621 caaagatggt gtggccgatg tgtctattga agattctgtg atctcactct caggagacca
     1681 ttgcatcatt ggccgcacac tggtggtcca tgaaaaagca gatgacttgg gcaaaggtgg
     1741 aaatgaagaa agtacaaaga caggaaacgc tggaagtcgt ttggcttgtg gtgtaattgg
     1801 gatcgcccaa gggtcgacgg taccgcgggc ccgggatcca ccggtcatgg tgagcaaggg
     1861 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     1921 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1981 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     2041 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     2101 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     2161 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     2221 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     2281 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     2341 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     2401 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     2461 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     2521 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtgagcgg ccgcgactct
     2581 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2641 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2821 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2881 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2941 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3001 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3061 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3121 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3181 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3241 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3301 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3361 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3421 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3481 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3541 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3601 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3661 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3721 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3781 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3961 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4321 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4561 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4621 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4681 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4741 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4801 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4861 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4921 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4981 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5041 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5101 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5161 gagcgtcaga ccccgtagaa aagatca
//
