LOCUS       sequence-337516-        6667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the purifcation of OPTN protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  AUTHORS   Chang C, Shi X, Jensen LE, Yokom AL, Fracchiolla D, Martens S, 
            Hurley JH
  TITLE     Reconstitution of cargo-induced LC3 lipidation in mammalian 
            selective autophagy.
  JOURNAL   Sci Adv. 2021 Apr 23;7(17). pii: 7/17/eabg4922. doi: 
            10.1126/sciadv.abg4922. Print 2021 Apr.
  PUBMED    33893090
REFERENCE   2  (bases 1 to 6667)
  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: "Sci Adv."; 
            date: "2021-04-23"; volume: "7(17). pii: 7/17/eabg4922. doi"; pages:
            " 10.1126/sciadv.abg4922. Print 2021 Apr"
COMMENT     Sequence Label: GST-OPTN
FEATURES             Location/Qualifiers
     source          1..6667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(148..153,154..171,172..178)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    186..202
                     /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     210..226
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     210..226
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             222..749
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(242..259)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(242..258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(251..273)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(851..856,857..872,873..879)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    887..903
                     /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)."
     CDS             926..1579
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1537..1559
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1903..1914
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3385..3407)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3552..3570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3638..3742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3743..3811,3812..4603)
                     /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(3961..3980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4774..5362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5263..5282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5516..5533
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5606..5683
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5684..6667,1..99)
                     /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(5703..5722)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
       61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
      121 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      181 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt
      241 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
      301 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
      361 gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac
      421 cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg
      481 tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtgacctatc
      541 ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca
      601 catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg
      661 ttggaattag cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg
      721 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
      781 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
      841 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
      901 caatttcaca caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg
      961 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt
     1021 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt
     1081 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat
     1141 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat
     1201 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag
     1261 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat
     1321 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga
     1381 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc
     1441 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta
     1501 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg
     1561 tggcgaccat cctccaaaaa tcgatatgtc ccatcaacct ctcagctgcc tcactgaaaa
     1621 ggaggacagc cccagtgaaa gcacaggaaa tggacccccc cacctggccc acccaaacct
     1681 ggacacgttt accccggagg agctgctgca gcagatgaaa gagctcctga ccgagaacca
     1741 ccagctgaaa gaagccatga agctaaataa tcaagccatg aaagggagat ttgaggagct
     1801 ttcggcctgg acagagaaac agaaggaaga acgccagttt tttgagatac agagcaaaga
     1861 agcaaaagag cgtctaatgg ccttgagtca tgagaatgag aaattgaagg aagagcttgg
     1921 aaaactaaaa gggaaatcag aaaggtcatc tgaggacccc actgatgact ccaggcttcc
     1981 cagggccgaa gcggagcagg aaaaggacca gctcaggacc caggtggtga ggctacaagc
     2041 agagaaggca gacctgttgg gcatcgtgtc tgaactgcag ctcaagctga actccagcgg
     2101 ctcctcagaa gattcctttg ttgaaattag gatggctgaa ggagaagcag aagggtcagt
     2161 aaaagaaatc aagcatagtc ctgggcccac gagaacagtc tccactggca cggcattgtc
     2221 taaatatagg agcagatctg cagatggggc caagaattac ttcgaacatg aggagttaac
     2281 tgtgagccag ctcctgctgt gcctaaggga agggaatcag aaggtggaga gacttgaagt
     2341 tgcactcaag gaggccaaag aaagagtttc agattttgaa aagaaaacaa gtaatcgttc
     2401 tgagattgaa acccagacag aggggagcac agagaaagag aatgatgaag agaaaggccc
     2461 ggagactgtt ggaagcgaag tggaagcact gaacctccag gtgacatctc tgtttaagga
     2521 gcttcaagag gctcatacaa aactcagcga agctgagcta atgaagaaga gacttcaaga
     2581 aaagtgtcag gcccttgaaa ggaaaaattc tgcaattcca tcagagttga atgaaaagca
     2641 agagcttgtt tatactaaca aaaagttaga gctacaagtg gaaagcatgc tatcagaaat
     2701 caaaatggaa caggctaaaa cagaggatga aaagtccaaa ttaactgtgc tacagatgac
     2761 acacaacaag cttcttcaag aacataataa tgcattgaaa acaattgagg aactaacaag
     2821 aaaagagtca gaaaaagtgg acagggcagt gctgaaggaa ctgagtgaaa aactggaact
     2881 ggcagagaag gctctggctt ccaaacagct gcaaatggat gaaatgaagc aaaccattgc
     2941 caagcaggaa gaggacctgg aaaccatgac catcctcagg gctcagatgg aagtttactg
     3001 ttctgatttt catgctgaaa gagcagcgag agagaaaatt catgaggaaa aggagcaact
     3061 ggcattgcag ctggcagttc tgctgaaaga gaatgatgct ttcgaagacg gaggcaggca
     3121 gtccttgatg gagatgcaga gtcgtcatgg ggcgagaaca agtgactctg accagcaggc
     3181 ttaccttgtt caaagaggag ctgaggacag ggactggcgg caacagcgga atattccgat
     3241 tcattcctgc cccaagtgtg gagaggttct gcctgacata gacacgttac agattcacgt
     3301 gatggattgc atcatttaac tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc
     3361 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3421 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3481 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt ataattcttg
     3541 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3601 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3961 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     4021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4261 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     4321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4921 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5401 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5461 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5521 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     5581 cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat
     5641 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     5701 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     5761 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     5821 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     5881 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     5941 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     6001 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     6061 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     6121 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     6181 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     6241 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     6301 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     6361 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     6421 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     6481 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     6541 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     6601 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     6661 aaaaacc
//
