LOCUS       Exported                5795 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Inducible expression of human alpha-synuclein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    pET21a-alpha-synuclein
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5795)
  TITLE     MJFF plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5795)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5795
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      20..475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(107..126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     317..338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        501..605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             606..1466
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(824..843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1637..2225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2126..2145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2379..2396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2411..2550
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2476..2495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2636..2658
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2652..2843)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3652..4734)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4696..4715
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4735..4812)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5018..5037
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5121..5140
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5121..5139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5140..5164
                     /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)."
     RBS             5179..5201
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5647..5664
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5717..5735)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5731..5778
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
       61 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      361 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      421 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
      481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1141 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2341 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2401 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2461 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     2521 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     2581 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2641 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     2701 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     2761 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     2821 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     2881 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     2941 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     3001 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     3061 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     3121 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     3181 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     3241 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     3301 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     3361 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     3421 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     3481 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     3541 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     3601 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     3661 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3721 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     3781 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     3841 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     3901 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     3961 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     4021 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     4081 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     4141 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     4201 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     4261 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     4321 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     4381 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     4441 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     4501 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     4561 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     4621 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     4681 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     4741 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     4801 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     4861 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     4921 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     4981 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5041 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     5101 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     5161 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat ggatgtgttt
     5221 atgaaaggtc tgagcaaagc gaaagaaggc gtggtggctg cggcggaaaa aacgaaacag
     5281 ggcgtggcgg aagcggccgg caaaacgaaa gaaggtgttc tgtatgtcgg cagcaaaacc
     5341 aaagaaggcg tggttcatgg tgtggccacc gttgcagaaa aaacgaaaga acaggtcacc
     5401 aacgtgggcg gtgctgtcgt gaccggtgtt acggctgtcg cgcaaaaaac ggtggaaggc
     5461 gcgggttcta ttgcggcggc aaccggtttc gttaaaaaag atcagctggg taaaaatgaa
     5521 gaaggcgcgc cgcaagaagg tatcctggaa gacatgccgg tggatccgga caacgaagcg
     5581 tatgaaatgc cgtcggaaga aggctatcaa gactatgaac cggaagcgta agcggccgca
     5641 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     5701 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     5761 gtcttgaggg gttttttgct gaaaggagga actat
//
