LOCUS       sequence-237743-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression construct for protein production and 
            purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Pastuzyn ED, Day CE, Kearns RB, Kyrke-Smith M, Taibi AV, McCormick 
            J, Yoder N, Belnap DM, Erlendsson S, Morado DR, Briggs JAG, 
            Feschotte C, Shepherd JD
  TITLE     The Neuronal Gene Arc Encodes a Repurposed Retrotransposon Gag 
            Protein that Mediates Intercellular RNA Transfer.
  JOURNAL   Cell. 2018 Mar 22;173(1):275. doi: 10.1016/j.cell.2018.03.024.
  PUBMED    29570995
REFERENCE   2  (bases 1 to 6157)
  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: "Cell."; 
            date: "2018-03-22"; volume: "173(1):275. doi"; pages: " 
            10.1016/j.cell.2018.03.024"
COMMENT     Sequence Label: pGEX-6p1-GST-ArcFL
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..654
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     612..634
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             661..684
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1950..1972)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2117..2135)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2203..2307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2308..2376,2377..3168)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2526..2545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3339..3927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3828..3847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4081..4098
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4171..4248
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4249..5331
                     /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     complement(4268..4287)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5380..5385,5386..5403,5404..5410)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5418..5434
                     /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     5442..5458
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5442..5458
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5454..5981
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5474..5491)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5474..5490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5483..5505)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6083..6088,6089..6104,6105..6111)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6119..6135
                     /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)."
ORIGIN
        1 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
       61 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
      121 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
      181 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
      241 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
      301 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
      361 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
      421 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
      481 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
      541 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
      601 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat
      661 ctggaagttc tgttccaggg gcccctggga tccatggagc tggaccatat gacgaccggc
      721 ggcctccacg cctaccctgc cccgcggggt gggccggccg ccaaacccaa tgtgatcctg
      781 cagattggta agtgccgagc tgagatgctg gagcacgtac ggaggaccca ccggcatctg
      841 ttgaccgaag tgtccaagca ggtggagcga gagctgaaag ggttgcacag gtcggtgggc
      901 aagctggaga acaacttgga cggctatgtg cccacgggcg actcacagcg ctggaagaag
      961 tccatcaagg cctgtctctg ccgctgccag gagaccatcg ccaacctgga gcgctgggtc
     1021 aagcgtgaga tgcacgtgtg gagggaggtc ttctaccgtc tggagaggtg ggccgaccgc
     1081 ctggagtcca tgggcggcaa gtacccagtg ggcagcgagc cggcccgcca cactgtctct
     1141 gtaggtgtgg ggggtccaga gccctactgc caggaagctg atggctacga ctacactgtt
     1201 agcccctatg ccatcacccc gccacctgcc gcaggagagc tgcctgagca ggagtcagtt
     1261 ggggctcagc aataccagtc ttgggtgcca ggtgaggatg ggcaaccaag cccaggtgtg
     1321 gatacccaga tctttgagga cccacgggag ttcctgagcc acctggaaga gtacctgcgg
     1381 caggtgggtg gctctgaaga atattggctg tcccagatcc agaaccacat gaatgggcca
     1441 gccaagaagt ggtgggagtt caaacagggc tcggtgaaga actgggtgga gttcaagaag
     1501 gagtttctgc agtacagtga gggtacgctc tcccgcgaag ccattcagcg ggagctggac
     1561 ctgccacaga agcagggtga gccacttgac cagttcctct ggcgtaagcg ggacctgtac
     1621 cagacactgt atgtggacgc tgaggaggag gagatcattc agtatgtggt gggcaccctg
     1681 cagcccaagt tcaagcgctt tctgcgccac ccacttccca agaccctgga gcagctcatc
     1741 cagaggggca tggaagttca ggacggcctg gagcaggcag ctgagccttc tgtcacccct
     1801 ctgcccacag aggatgagac tgaggcactc acgcctgctc ttaccagcga gtcagtagcc
     1861 agtgacagga cccagcctga atagctcgag cggccgcatc gtgactgact gacgatctgc
     1921 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1981 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2041 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat
     2101 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     2161 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2521 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2821 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     2881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3181 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3241 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3301 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3421 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3481 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3541 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3601 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3661 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3721 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3781 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3841 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3901 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3961 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4021 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4081 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     4141 ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggcgcaa aacctttcgc
     4201 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     4261 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     4321 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     4381 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     4441 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     4501 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     4561 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     4621 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     4681 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     4741 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     4801 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     4861 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     4921 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     4981 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     5041 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     5101 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     5161 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     5221 aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5281 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5341 aattaatgta agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5401 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5461 tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5521 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     5581 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct
     5641 ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg
     5701 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
     5761 acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt
     5821 gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta
     5881 tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct tctggcgtca
     5941 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     6001 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     6061 gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt
     6121 gtgagcggat aacaatttca cacaggaaac agtattc
//
