LOCUS       sequence-361481-        6634 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 6634)
  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 6634)
  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
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             663..692
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1237..1254)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2857..2876)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3304..3324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3309..3638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3489..3624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3676..3695)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3677..3695)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3709..3725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3709..3725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3722..3744)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3733..3749
                     /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(3757..3763,3764..3781,3782..3787))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3802..3823
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3940..3957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4111..4699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4191..4210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4870..5661,5662..5730))
                     /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     5493..5512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5731..5835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5862..6317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5949..5968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6159..6180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6443..6465
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6457..6474
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6458..6474
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaatt cagatctggt accacgcgta tcgataagct tgcatgcctg caggtcgaca
      661 tggaacagaa actcatctct gaagaggatc tgtctagagg atccatgtca aataacggcc
      721 tagacattca agacaaaccc ccagcccctc cgatgagaaa taccagcact atgattggag
      781 ccggcagcaa agatgctgga accctaaacc atggttctaa acctctgcct ccaaacccag
      841 aggagaagaa aaagaaggac cgattttacc gatccatttt acctggagat aaaacaaata
      901 aaaagaaaga gaaagagcgg ccagagattt ctctcccttc agattttgaa cacacaattc
      961 atgtcggttt tgatgctgtc acaggggagt ttacgggaat gccagagcag tgggcccgct
     1021 tgcttcagac atcaaatatc actaagtcgg agcagaagaa aaacccgcag gctgttctgg
     1081 atgtgttgga gttttacaac tcgaagaaga catccaacag ccagaaatac atgagcttta
     1141 cagataagtc agctgaggat tacaattctt ctaatgcctt gaatgtgaag gctgtgtctg
     1201 agactcctgc agtgccacca gtttcagaag atgaggatga tgatgatgat gatgctaccc
     1261 caccaccagt gattgctcca cgcccagagc acacaaaatc tgtatacaca cggtctgtga
     1321 ttgaaccact tcctgtcact ccaactcggg acgtggctac atctcccatt tcacctactg
     1381 aaaataacac cactccacca gatgctttga cccggaatac tgagaagcag aagaagaagc
     1441 ctaaaatgtc tgatgaggag atcttggaga aattacgaag catagtgagt gtgggcgatc
     1501 ctaagaagaa atatacacgg tttgagaaga ttggacaagg tgcttcaggc accgtgtaca
     1561 cagcaatgga tgtggccaca ggacaggagg tggccattaa gcagatgaat cttcagcagc
     1621 agcccaagaa agagctgatt attaatgaga tcctggtcat gagggaaaac aagaacccaa
     1681 acattgtgaa ttacttggac agttacctcg tgggagatga gctgtgggtt gttatggaat
     1741 acttggctgg aggctccttg acagatgtgg tgacagaaac ttgcatggat gaaggccaaa
     1801 ttgcagctgt gtgccgtgag tgtctgcagg ctctggagtt cttgcattcg aaccaggtca
     1861 ttcacagaga catcaagagt gacaatattc tgttgggaat ggatagctct gtcaagctaa
     1921 ctgactttgg attctgtgca cagataaccc cagagcagag caaacggagc accatggtag
     1981 gaaccccata ctggatggca ccagaggttg tgacacgaaa ggcctatggg cccaaggttg
     2041 acatctggtc cctgggcatc atggccatcg aaatgattga aggggagcct ccatacctca
     2101 atgaaaaccc tctgagagcc ttgtacctca ttgccaccaa tgggacccca gaacttcaga
     2161 acccagagaa gctgtcagct atcttccggg actttctgaa ccgctgtctc gagatggatg
     2221 tggagaagag aggttcagct aaagagctga tacagcatca attcctgaag attgccaagc
     2281 ccctctccag cctcactcca ctgattgctg cagctaagga ggcaacaaag aacaatcact
     2341 aaaaccacac tcaccccagc ctcattgtgc caagccttct gtgagataaa tgcacatttc
     2401 agaaattcca actcctgatg ccctcttctc cttgccttgc ttctcccatt tcctgatcta
     2461 gcactcctca agactttgat ccttggaaac cgtgtgtcca gcattgaaga gaactgcaac
     2521 tgaatgacta atcagatgat ggccatttct aaataaggaa tttcctccca attcatggat
     2581 atgagggtgg tttatgatta agggtttata taaataaatg tttctagtct tccgggcctt
     2641 cgtggcctcg aggaattccg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     2701 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     2761 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     2821 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     2881 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     2941 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     3001 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     3061 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     3121 ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc
     3181 ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc
     3241 tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc
     3301 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3361 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3541 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3601 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcctcgag gaactgaaaa
     3661 accagaaagt taattcccta tagtgagtcg tattaaattc gtaatcatgt catagctgtt
     3721 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3781 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3841 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3901 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     5881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6001 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6061 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6181 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6301 attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6361 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     6421 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     6481 ctgatctata cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca
     6541 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     6601 tatattggct catgtccaat atgaccgcca tgtt
//
