LOCUS       sequence-413253-        6397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6397)
  TITLE     GSK3 plasmids
REFERENCE   2  (bases 1 to 6397)
  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"
COMMENT     Sequence Label: GSK3 beta pGEX
FEATURES             Location/Qualifiers
     source          1..6397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1574..1585
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2144..2160)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2145..2161)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2229..2251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2396..2414)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2482..2586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2587..2655,2656..3447)
                     /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(2805..2824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3618..4206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4107..4126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4360..4377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4450..4527
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4528..5610
                     /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(4547..4566)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5659..5664,5665..5682,5683..5689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5697..5713
                     /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     5721..5737
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5721..5737
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5733..6260
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5753..5770)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5753..5769)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5762..5784)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6362..6367,6368..6383,6384..6390)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccccg
      721 ggaatttccg gtggtggtgg tggaattcta gactccatgg cggggcgacc gagaaccacc
      781 tcctttgcgg agagctgcaa gccagtgcag cagccttcag cttttggtag catgaaagtt
      841 agcagagata aagatggcag caaggtaacc acagtggtgg caactcctgg acagggtcct
      901 gacaggccac aggaagtcag ttacacagac actaaagtca ttggaaatgg gtcatttggt
      961 gtggtatatc aagccaaact ttgtgactca ggagaactgg tggccatcaa gaaagttctt
     1021 caggacaagc gatttaagaa ccgagagctc cagatcatga gaaagctaga tcactgtaac
     1081 atagtccgat tgcggtattt cttctactcg agtggcgaga agaaagatga ggtctacctt
     1141 aacctggtgc tggactatgt tccggaaaca gtgtacagag tcgccagaca ctatagtcga
     1201 gccaagcaga cactccctgt gatctatgtc aagttgtata tgtaccagct gttcagaagt
     1261 ctagcctata tccattcctt tgggatctgc catcgagaca ttaaaccaca gaacctcttg
     1321 ctggatcctg atactgctgt attaaaactc tgtgactttg gaagtgcaaa gcagctggtc
     1381 cgaggagaac ccaatgtttc gtatatctgt tctcggtact atagggcacc agagttgatc
     1441 tttggagcca ctgattatac ctctagtata gatgtatggt ctgctggctg tgtgttggct
     1501 gagctgttac taggacaacc aatatttcca ggggatagtg gtgtggatca gttggtagaa
     1561 ataatcaagg tcctgggaac tccaacaagg gagcaaatca gagaaatgaa cccaaactac
     1621 acagaattta aattccctca aattaaggca catccttgga ctaaggtctt ccgaccccga
     1681 actccaccgg aggcaattgc actgtgtagc cgtctgctgg agtatacacc aactgcccga
     1741 ctaacaccac tggaagcttg tgcacattca ttttttgatg aattacggga cccaaatgtc
     1801 aaactaccaa atgggcgaga cacacctgca ctcttcaact tcaccactca agaactgtca
     1861 agtaatccac ctctggctac catccttatt cctcctcatg ctcggattca agcagctgct
     1921 tcaaccccca caaatgccac agcagcgtca gatgctaata ctggagaccg tggacagacc
     1981 aataatgctg cttctgcatc agcttccaac tccacctgaa cagtcccgag cagccagctg
     2041 cacaggaaaa accaccagtt acttgagtgt cactcagcaa cactggtcac gtttggaaag
     2101 aatattaaaa aaaaaaaaaa aaggaattcg atatcaagct tatcgatacc gtcgacctcg
     2161 agctcaagct taattcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac
     2221 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2281 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     2341 gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc
     2401 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2461 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2521 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2581 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2641 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2701 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2761 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2821 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2881 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2941 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3001 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3061 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3121 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3181 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3241 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3301 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3361 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3421 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3481 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3541 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3601 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3661 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3721 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3781 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3841 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3901 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3961 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4021 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4081 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4141 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4201 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4261 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4321 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4381 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4441 ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     4501 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     4561 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     4621 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     4681 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     4741 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     4801 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     4861 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     4921 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     4981 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     5041 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     5101 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     5161 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     5221 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     5281 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     5341 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     5401 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     5461 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     5521 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5581 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     5641 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5701 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg
     5761 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5821 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5881 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg
     5941 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa
     6001 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg
     6061 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg
     6121 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta
     6181 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     6241 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     6301 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     6361 gttgacaatt aatcatcggc tcgtataatg tgtggaa
//
