LOCUS       Exported                5924 bp ds-DNA     circular SYN 16-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pF155 pcDNA3.1(+)SOD1G93A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  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 5924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5924
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        28..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     656..675
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        656..674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1256..1273)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1262..1486
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1532..1960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1619..1638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1829..1850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1969..1989)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1974..2303
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2154..2289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2216..2235
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2370..3164
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2424..2443)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3034..3053
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3338..3459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3375..3394)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3429..3448
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3508..3524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3508..3524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3521..3543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3532..3548
                     /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)."
     promoter        complement(3556..3586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3601..3622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3791..3808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3962..4550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4042..4061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4721..5581)
                     /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     5344..5363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5582..5686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5761..5780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      661 cgactcacta tagggagacc caagctggct agcgtttaaa cttaagcttg gtaccgagct
      721 cggatccact agtccagtgt ggtggaattc catggcgacg aaggccgtgt gcgtgctgaa
      781 gggcgacggc ccagtgcagg gcatcatcaa tttcgagcag aaggaaagta atggaccagt
      841 gaaggtgtgg ggaagcatta aaggactgac tgaaggcctg catggattcc atgttcatga
      901 gtttggagat aatacagcag gctgtaccag tgcaggtcct cactttaatc ctctatccag
      961 aaaacacggt gggccaaagg atgaagagag gcatgttgga gacttgggca atgtgactgc
     1021 tgacaaagat gctgtggccg atgtgtctat tgaagattct gtgatctcac tctcaggaga
     1081 ccattgcatc attggccgca cactggtggt ccatgaaaaa gcagatgact tgggcaaagg
     1141 tggaaatgaa gaaagtacaa agacaggaaa cgctggaagt cgtttggctt gtggtgtaat
     1201 tgggatcgcc caataacgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     1261 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1321 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     1381 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     1441 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     1501 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     1561 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1621 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1681 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1741 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1801 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1861 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1921 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     1981 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2041 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2101 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2161 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2221 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     2281 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     2341 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2401 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2461 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2521 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     2581 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2641 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2701 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2761 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2821 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2881 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2941 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3001 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3061 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3121 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3181 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3241 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3301 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     3361 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3421 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     3481 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3721 tgggcgctca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3781 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3841 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3901 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4381 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4621 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4681 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4801 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4861 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4921 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4981 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5041 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5101 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5161 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5221 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5281 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5341 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5401 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5461 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5521 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5581 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5641 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5701 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     5821 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     5881 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttc
//
