LOCUS       sequence-359588-        5197 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 5197)
  AUTHORS   Kim SH, Li Z, Sacks DB
  TITLE     E-cadherin-mediated cell-cell attachment activates Cdc42.
  JOURNAL   J Biol Chem. 2000 Nov 24. 275(47):36999-7005.
  PUBMED    10950951
REFERENCE   2  (bases 1 to 5197)
  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: "J Biol 
            Chem. 2000 Nov 24. 275(47):36999-7005."
COMMENT     Sequence Label: pGEX-KG-WASP-GBD
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..82
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        155..232
                     /gene="lacI"
                     /label=lacI promoter
     CDS             233..1315
                     /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(252..271)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1364..1369,1370..1387,1388..1394)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1402..1418
                     /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     1426..1442
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1426..1442
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1438..1965
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1458..1475)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1458..1474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1467..1489)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2067..2072,2073..2088,2089..2095)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2103..2119
                     /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             2142..2795
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2753..2775
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2802..2819
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3131..3153)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3298..3316)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3384..3488
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3489..3557,3558..4349)
                     /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(3707..3726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4520..5108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5009..5028
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
       61 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
      121 gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc gcaaaacctt
      181 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
      241 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
      301 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
      361 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
      421 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
      481 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
      541 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
      601 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
      661 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
      721 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
      781 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
      841 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
      901 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
      961 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     1021 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     1081 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     1141 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     1201 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1261 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1321 acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1381 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1441 accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1501 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1561 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt
     1621 gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag
     1681 gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac
     1741 accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg
     1801 ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga
     1861 attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa tgcttctggc
     1921 gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc
     1981 gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt
     2041 tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg
     2101 aattgtgagc ggataacaat ttcacacagg aaacagtatt catgtcccct atactaggtt
     2161 attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa
     2221 aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg
     2281 aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac
     2341 agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa
     2401 aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt
     2461 cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac
     2521 ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc
     2581 atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc
     2641 caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc
     2701 cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc
     2761 aagccacgtt tggtggtggc gaccatcctc caaaatcgga tctggttccg cgtggatcct
     2821 cagggaagaa gaagatcagc aaagctgata ttggtgcacc cagtggattc aagcatgtca
     2881 gccacgtggg gtgggacccc cagaatggat ttgacgtgaa caacctcgac ccagatctgc
     2941 ggagtctgtt ctccagggca ggaatcagcg aggcccagct caccgacgcc gagacctcta
     3001 aacttatcta cgacttcatt gaggaccagg gttaagaatt ctagactcca tgggtcgact
     3061 cgagctcaag cttaattcat cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg
     3121 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     3181 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     3241 cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc
     3301 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3481 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3601 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3661 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3721 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3781 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3901 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4021 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4081 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4321 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4501 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4561 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4621 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4681 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4741 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4801 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4861 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4921 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4981 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5041 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5101 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5161 ctcacatgtt ctttcctgcg ttatcccctg attctgt
//
