LOCUS       Exported                7921 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    plv-AcGFP-SOD1 (wt)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7921)
  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 7921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7921
                     /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"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      595..604
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             601..1317
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    1825..2413
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1878..1898)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2296..2307)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2500..2733
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2805..2926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2842..2861)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2896..2915
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2966..3101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3028..3047
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3121..3140)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3122..3140)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3150..3167)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3150..3166)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3159..3181)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3308..3763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3395..3414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3605..3626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3789..3893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3894..4754
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4112..4131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4925..5513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5414..5433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5667..5684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5801..5822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5837..5867
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5875..5891
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5880..5902
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5899..5915
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5899..5915
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5934..5954
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5936..5954
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5982..6208
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6209..6389
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6436..6561
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7054..7287
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7472..7516
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7783..7899
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
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 gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgactc tagcgctacc ggtcgccacc
      601 atggtgagca agggcgccga gctgttcacc ggcatcgtgc ccatcctgat cgagctgaat
      661 ggcgatgtga atggccacaa gttcagcgtg agcggcgagg gcgagggcga tgccacctac
      721 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcctgtgcc ctggcccacc
      781 ctggtgacca ccctgagcta cggcgtgcag tgcttctcac gctaccccga tcacatgaag
      841 cagcacgact tcttcaagag cgccatgcct gagggctaca tccaggagcg caccatcttc
      901 ttcgaggatg acggcaacta caagtcgcgc gccgaggtga agttcgaggg cgataccctg
      961 gtgaatcgca tcgagctgac cggcaccgat ttcaaggagg atggcaacat cctgggcaat
     1021 aagatggagt acaactacaa cgcccacaat gtgtacatca tgaccgacaa ggccaagaat
     1081 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg atggcagcgt gcagctggcc
     1141 gaccactacc agcagaatac ccccatcggc gatggccctg tgctgctgcc cgataaccac
     1201 tacctgtcca cccagagcgc cctgtccaag gaccccaacg agaagcgcga tcacatgatc
     1261 tacttcggct tcgtgaccgc cgccgccatc acccacggca tggatgagct gtacaagtcc
     1321 ggactcagat ctcgagctca agcttcgaat tccatggcga cgaaggccgt gtgcgtgctg
     1381 aagggcgacg gcccagtgca gggcatcatc aatttcgagc agaaggaaag taatggacca
     1441 gtgaaggtgt ggggaagcat taaaggactg actgaaggcc tgcatggatt ccatgttcat
     1501 gagtttggag ataatacagc aggctgtacc agtgcaggtc ctcactttaa tcctctatcc
     1561 agaaaacacg gtgggccaaa ggatgaagag aggcatgttg gagacttggg caatgtgact
     1621 gctgacaaag atggtgtggc cgatgtgtct attgaagatt ctgtgatctc actctcagga
     1681 gaccattgca tcattggccg cacactggtg gtccatgaaa aagcagatga cttgggcaaa
     1741 ggtggaaatg aagaaagtac aaagacagga aacgctggaa gtcgtttggc ttgtggtgta
     1801 attgggatcg cccaataagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     1861 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1921 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1981 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2041 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2101 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2161 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2221 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2281 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2341 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2401 gccgcctccc cgcctggaat tcgagctcgg tacctttaag accaatgact tacaaggcag
     2461 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     2521 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     2581 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     2641 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     2701 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     2761 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     2821 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2881 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     2941 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     3121 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     3181 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3241 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3301 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3361 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3601 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3661 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3721 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     3781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4861 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5041 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5101 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5161 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5221 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5281 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5341 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5401 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5461 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5521 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5581 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5641 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5701 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5761 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     5821 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     5881 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     5941 accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact
     6001 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
     6061 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
     6121 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
     6181 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
     6241 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6301 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6361 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     6421 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     6481 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     6541 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     6601 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     6661 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     6721 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     6781 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     6841 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     6901 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     6961 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     7021 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     7081 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     7141 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     7201 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     7261 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     7321 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     7381 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     7441 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     7501 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     7561 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     7621 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     7681 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
     7741 atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa gaaaaggggg
     7801 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     7861 taaagaatta caaaaacaaa ttacaaaatt caaaatttta tcgataagct tgggagttcc
     7921 g
//
