LOCUS       sequence-158653-        6699 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 6699)
  AUTHORS   Sells MA, Knaus UG, Bagrodia S, Ambrose DM, Bokoch GM, Chernoff J
  TITLE     Human p21-activated kinase (Pak1) regulates actin organization in 
            mammalian cells.
  JOURNAL   Curr Biol. 1997 Mar 1. 7(3):202-10.
  PUBMED    9395435
REFERENCE   2  (bases 1 to 6699)
  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: "Curr Biol. 
            1997 Mar 1. 7(3):202-10."
COMMENT     Sequence Label: pCMV6M-Pak1 H83L H86L
FEATURES             Location/Qualifiers
     source          1..6699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2580..2597
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2581..2597
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2758..3137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3138..3341
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3292..3312
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3413..3442
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3987..4004)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5607..5626)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5982..6002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6144..6460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6311..6446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6498..6517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6499..6517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6531..6547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6531..6547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6544..6566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6555..6571
                     /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)."
     promoter        complement(join(6579..6585,6586..6603,6604..6609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6624..6645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2521 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2581 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
     2641 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     2701 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     2761 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2821 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2881 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2941 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3001 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3061 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     3121 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     3181 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3241 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     3301 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     3361 attaattcag atctggtacc acgcgtatcg ataagcttcg atcgtcgaca tggaacagaa
     3421 actcatctct gaagaggatc tgtctagagg atccatgtca aataacggcc tagacattca
     3481 agacaaaccc ccagcccctc cgatgagaaa taccagcact atgattggag ccggcagcaa
     3541 agatgctgga accctaaacc atggttctaa acctctgcct ccaaacccag aggagaagaa
     3601 aaagaaggac cgattttacc gatccatttt acctggagat aaaacaaata aaaagaaaga
     3661 gaaagagcgg ccagagattt ctctcccttc agattttgag ctcacaattc ttgtcggttt
     3721 tgatgctgtc acaggggagt ttacgggaat gccagagcag tgggcccgct tgcttcagac
     3781 atcaaatatc actaagtcgg agcagaagaa aaacccgcag gctgttctgg atgtgttgga
     3841 gttttacaac tcgaagaaga catccaacag ccagaaatac atgagcttta cagataagtc
     3901 agctgaggat tacaattctt ctaatgcctt gaatgtgaag gctgtgtctg agactcctgc
     3961 agtgccacca gtttcagaag atgaggatga tgatgatgat gatgctaccc caccaccagt
     4021 gattgctcca cgcccagagc acacaaaatc tgtatacaca cggtctgtga ttgaaccact
     4081 tcctgtcact ccaactcggg acgtggctac atctcccatt tcacctactg aaaataacac
     4141 cactccacca gatgctttga cccggaatac tgagaagcag aagaagaagc ctaaaatgtc
     4201 tgatgaggag atcttggaga aattacgaag catagtgagt gtgggcgatc ctaagaagaa
     4261 atatacacgg tttgagaaga ttggacaagg tgcttcaggc accgtgtaca cagcaatgga
     4321 tgtggccaca ggacaggagg tggccattaa gcagatgaat cttcagcagc agcccaagaa
     4381 agagctgatt attaatgaga tcctggtcat gagggaaaac aagaacccaa acattgtgaa
     4441 ttacttggac agttacctcg tgggagatga gctgtgggtt gttatggaat acttggctgg
     4501 aggctccttg acagatgtgg tgacagaaac ttgcatggat gaaggccaaa ttgcagctgt
     4561 gtgccgtgag tgtctgcagg ctctggagtt cttgcattcg aaccaggtca ttcacagaga
     4621 catcaagagt gacaatattc tgttgggaat ggatagctct gtcaagctaa ctgactttgg
     4681 attctgtgca cagataaccc cagagcagag caaacggagc accatggtag gaaccccata
     4741 ctggatggca ccagaggttg tgacacgaaa ggcctatggg cccaaggttg acatctggtc
     4801 cctgggcatc atggccatcg aaatgattga aggggagcct ccatacctca atgaaaaccc
     4861 tctgagagcc ttgtacctca ttgccaccaa tgggacccca gaacttcaga acccagagaa
     4921 gctgtcagct atcttccggg actttctgaa ccgctgtctc gagatggatg tggagaagag
     4981 aggttcagct aaagagctga tacagcatca attcctgaag attgccaagc ccctctccag
     5041 cctcactcca ctgattgctg cagctaagga ggcaacaaag aacaatcact aaaaccacac
     5101 tcaccccagc ctcattgtgc caagccttct gtgagataaa tgcacatttc agaaattcca
     5161 actcctgatg ccctcttctc cttgccttgc ttctcccatt tcctgatcta gcactcctca
     5221 agactttgat ccttggaaac cgtgtgtcca gcattgaaga gaactgcaac tgaatgacta
     5281 atcagatgat ggccatttct aaataaggaa tttcctccca attcatggat atgagggtgg
     5341 tttatgatta agggtttata taaataaatg tttctagtct tccgggcctt cgtggcctcg
     5401 aggaattccg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     5461 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     5521 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     5581 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     5641 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     5701 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     5761 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     5821 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     5881 ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc
     5941 tggccatgcc caaccggtgg gacatttgag ttgcttgctg tggaatgtgt gtcagttagg
     6001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6061 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6121 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6181 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     6481 aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     6541 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6601 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctg
//
