LOCUS       sequence-435646-        6258 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 6258)
  AUTHORS   Xiao GH, Beeser A, Chernoff J, Testa JR
  TITLE     p21-activated kinase links Rac/Cdc42 signaling to merlin.
  JOURNAL   J Biol Chem. 2002 Jan 11. 277(2):883-6.
  PUBMED    11719502
REFERENCE   2  (bases 1 to 6258)
  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. 2002 Jan 11. 277(2):883-6."
COMMENT     Sequence Label: pEBG-Pak1 83-149
FEATURES             Location/Qualifiers
     source          1..6258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    112..133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     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     191..213
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     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"
     promoter        258..468
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        280..476
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      327..462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        563..1741
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          794..1732
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1689..1709
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1793..2446
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2404..2426
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2453..2470
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3392..3409)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3392..3408)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3401..3423)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3621..4076
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3708..3727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3918..3939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4093..4112)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4212..4234
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4272..4290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4358..4462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4463..4531,4532..5323)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4681..4700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5494..6082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5983..6002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6236..6253
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
       61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      121 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      181 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      241 gcttaattcc ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      301 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      361 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
      421 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
      481 tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
      541 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      601 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      661 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      721 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      781 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      841 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
      901 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
      961 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     1021 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     1081 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     1141 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     1201 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     1261 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     1321 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     1381 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     1441 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1501 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     1561 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     1621 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     1681 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     1741 aggaattctc tagagatccc tcgacctcga gatccattgt gctggatcta caatggcccc
     1801 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata
     1861 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa
     1921 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt
     1981 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg
     2041 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag
     2101 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct
     2161 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt
     2221 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt
     2281 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat
     2341 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt
     2401 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggttcc
     2461 gcgtggatcc cacacaattc atgtcggttt tgatgctgtc acaggggagt ttacgggaat
     2521 gccagagcag tgggcccgct tgcttcagac atcaaatatc actaagtcgg agcagaagaa
     2581 aaacccgcag gctgttctgg atgtgttgga gttttacaac tcgaagaaga catccaacag
     2641 ccagaaatac atgagcttta cagataagtc atgaattcgg taccatcgat gcggccgcga
     2701 ctctagagtg agggtcccca cctgggaccc ttgagagtat caggtctccc acgtgggaga
     2761 caagaaatcc ctgtttaata tttaaacagc agtgttcccc atctgggtcc ttgcacccct
     2821 cactctggcc tcagccgact gcacagcggc ccctgcatcc ccttggctgt gaggcccctg
     2881 gacaagcaga ggtggccaga gctgggaggc atggccctgg ggtcccacga atttgctggg
     2941 gaatctcgtt tttcttctta agacttttgg gacatggttt gactcccgaa catcaccgac
     3001 gtgtctcctg tttttctggg tggcctcggg acacctgccc tgcccccacg agggtcagga
     3061 ctgtgactct ttttagggcc aggcaggtgc ctggacattt gccttgctgg acggggactg
     3121 gggatgtggg agggagcaga caggaggaat catgtcaggc ctgtgtgtga aaggaagctc
     3181 cactgtcacc ctccacctct tcacccccca ctcaccagtg tcccctccac tgtcacattg
     3241 taactgaact tcaggataat aaagtgtttg cctccaaaaa aaaaaaaaaa aaaaaaaaaa
     3301 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3361 aaaaaaaaaa aaaaaaaaaa aaaccgaatt cactggccgt cgttttacaa cgtcgtgact
     3421 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3481 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3541 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3601 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3661 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3721 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3781 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3841 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3901 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     3961 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4021 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4081 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4141 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4201 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4261 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4321 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4381 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4441 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4501 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4561 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4621 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4681 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4741 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4801 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4861 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4921 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4981 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5041 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5101 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5161 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5221 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5281 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5641 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6181 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6241 gtcagtgagc gaggaagc
//
