LOCUS       sequence-223090-        6627 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 6627)
  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 6627)
  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 K299R
FEATURES             Location/Qualifiers
     source          1..6627
                     /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             656..685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1230..1247)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2850..2869)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3297..3317)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3302..3631
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3482..3617
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3669..3688)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3670..3688)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3702..3718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3702..3718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3715..3737)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3726..3742
                     /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(3750..3756,3757..3774,3775..3780))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3795..3816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3933..3950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4104..4692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4184..4203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4863..5654,5655..5723))
                     /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     5486..5505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5724..5828)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5855..6310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5942..5961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6152..6173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6436..6458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6450..6467
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6451..6467
                     /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 tcgatcgtcg acatggaaca
      661 gaaactcatc tctgaagagg atctgtctag aggatccatg tcaaataacg gcctagacat
      721 tcaagacaaa cccccagccc ctccgatgag aaataccagc actatgattg gagccggcag
      781 caaagatgct ggaaccctaa accatggttc taaacctctg cctccaaacc cagaggagaa
      841 gaaaaagaag gaccgatttt accgatccat tttacctgga gataaaacaa ataaaaagaa
      901 agagaaagag cggccagaga tttctctccc ttcagatttt gaacacacaa ttcatgtcgg
      961 ttttgatgct gtcacagggg agtttacggg aatgccagag cagtgggccc gcttgcttca
     1021 gacatcaaat atcactaagt cggagcagaa gaaaaacccg caggctgttc tggatgtgtt
     1081 ggagttttac aactcgaaga agacatccaa cagccagaaa tacatgagct ttacagataa
     1141 gtcagctgag gattacaatt cttctaatgc cttgaatgtg aaggctgtgt ctgagactcc
     1201 tgcagtgcca ccagtttcag aagatgagga tgatgatgat gatgatgcta ccccaccacc
     1261 agtgattgct ccacgcccag agcacacaaa atctgtatac acacggtctg tgattgaacc
     1321 acttcctgtc actccaactc gggacgtggc tacatctccc atttcaccta ctgaaaataa
     1381 caccactcca ccagatgctt tgacccggaa tactgagaag cagaagaaga agcctaaaat
     1441 gtctgatgag gagatcttgg agaaattacg aagcatagtg agtgtgggcg atcctaagaa
     1501 gaaatataca cggtttgaga agattggaca aggtgcttca ggcaccgtgt acacagcaat
     1561 ggatgtggcc acaggacagg aggtcgcgat taggcagatg aatcttcagc agcagcccaa
     1621 gaaagagctg attattaatg agatcctggt catgagggaa aacaagaacc caaacattgt
     1681 gaattacttg gacagttacc tcgtgggaga tgagctgtgg gttgttatgg aatacttggc
     1741 tggaggctcc ttgacagatg tggtgacaga aacttgcatg gatgaaggcc aaattgcagc
     1801 tgtgtgccgt gagtgtctgc aggctctgga gttcttgcat tcgaaccagg tcattcacag
     1861 agacatcaag agtgacaata ttctgttggg aatggatggc tctgtcaagc taactgactt
     1921 tggattctgt gcacagataa ccccagagca gagcaaacgg agcaccatgg taggaacccc
     1981 atactggatg gcaccagagg ttgtgacacg aaaggcctat gggcccaagg ttgacatctg
     2041 gtccctgggc atcatggcca tcgaaatgat tgaaggggag cctccatacc tcaatgaaaa
     2101 ccctctgaga gccttgtacc tcattgccac caatgggacc ccagaacttc agaacccaga
     2161 gaagctgtca gctatcttcc gggactttct gaaccgctgt ctcgagatgg atgtggagaa
     2221 gagaggttca gctaaagagc tgatacagca tcaattcctg aagattgcca agcccctctc
     2281 cagcctcact ccactgattg ctgcagctaa ggaggcaaca aagaacaatc actaaaacca
     2341 cactcacccc agcctcattg tgccaagcct tctgtgagat aaatgcacat ttcagaaatt
     2401 ccaactcctg atgccctctt ctccttgcct tgcttctccc atttcctgat ctagcactcc
     2461 tcaagacttt gatccttgga aaccgtgtgt ccagcattga agagaactgc aactgaatga
     2521 ctaatcagat gatggccatt tctaaataag gaatttcctc ccaattcatg gatatgaggg
     2581 tggtttatga ttaagggttt atataaataa atgtttctag tcttccgggc cttcgtggcc
     2641 tcgaggaatt ccgtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     2701 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2761 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2821 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     2881 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     2941 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     3001 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3061 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3121 tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct
     3181 acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg
     3241 cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa
     3301 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3361 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3421 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3481 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3541 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3601 aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa
     3661 agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg
     3721 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3781 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4501 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5821 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     5881 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5941 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6001 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6061 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     6241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     6301 cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6361 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6421 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct
     6481 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
     6541 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     6601 gctcatgtcc aatatgaccg ccatgtt
//
