LOCUS       sequence-252938-        5960 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 5960)
  AUTHORS   Ho HY, Rohatgi R, Lebensohn AM, Le Ma, Li J, Gygi SP, Kirschner MW
  TITLE     Toca-1 mediates Cdc42-dependent actin nucleation by activating the 
            N-WASP-WIP complex.
  JOURNAL   Cell. 2004 Jul 23;118(2):203-16.
  PUBMED    15260990
REFERENCE   2  (bases 1 to 5960)
  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: "Cell."; 
            date: "2004-07-23"; volume: "118(2)"; pages: "203-16"
COMMENT     Sequence Label: pCS2+MT-hToca-1-wt
FEATURES             Location/Qualifiers
     source          1..5960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        71..1055
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1090..1108
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1090..1107
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1158..1187
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1197..1226
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1236..1265
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1275..1304
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1338..1367
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3067..3201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3091..3110)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3145..3164
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3314..3334)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3314..3332)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3353..3369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3353..3369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3366..3388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3377..3393
                     /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(3401..3407,3408..3425,3426..3431))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3446..3467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3584..3601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3755..4343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3835..3854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4514..5305,5306..5374))
                     /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     5137..5156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5375..5479)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5505..5960)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5642..5663)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5854..5873
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
      121 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
      181 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
      241 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
      301 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
      361 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
      421 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
      481 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
      541 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
      601 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
      661 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
      721 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
      781 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
      841 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
      901 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
      961 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     1021 gggaggtcta tataagcaat gctcgtttag ggaaccgcca ttctgcctgg ggacgtcgga
     1081 gcaagcttga tttaggtgac actatagaat acaagctact tgttcttttt gcaggatccc
     1141 atcgatttaa agctatggag caaaagctca tttctgaaga ggacttgaat gaaatggagc
     1201 aaaagctcat ttctgaagag gacttgaatg aaatggagca aaagctcatt tctgaagagg
     1261 acttgaatga aatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagagct
     1321 tgggcgacct caccatggag caaaagctca tttctgaaga ggacttgaat tcaggccggc
     1381 ctaccatgag ctggggcacg gagctgtggg atcagttcga cagcttagac aagcatacac
     1441 aatggggaat tgacttcttg gaaagatatg ccaaatttgt taaagagagg atagaaattg
     1501 aacagaacta tgcgaaacaa ttgagaaatc tggttaagaa gtactgcccc aaacgttcat
     1561 ccaaagatga agagccacgg tttacctcgt gtgtagcctt ttttaatatc cttaatgagt
     1621 taaatgacta tgcaggacag cgagaagttg tagcagaaga aatggcgcac agagtgtatg
     1681 gtgaattaat gagatatgct catgatctga aaactgaaag aaaaatgcat ctgcaagaag
     1741 gacgaaaagc tcaacaatat ctggatatgt gctggaaaca gatggataat agtaaaaaga
     1801 agtttgaaag agaatgtaga gaggcagaaa aggcacaaca gagttatgaa agattggata
     1861 atgatactaa tgcaaccaag gcagatgttg aaaaggccaa acagcagttg aatctgcgta
     1921 cgcatatggc cgatgaaaat aaaaatgaat atgctgcaca attacaaaac tttaatggag
     1981 aacaacataa acatttttat gtagtgattc ctcagattta caagcaacta caagaaatgg
     2041 acgaacgaag gactattaaa ctcagtgagt gttacagagg atttgctgac tcagaacgca
     2101 aagttattcc catcatttca aaatgtttgg aaggaatgat tcttgcagca aaatcagttg
     2161 atgaaagaag agactctcaa atggtggtag actccttcaa atctggtttt gaacctccag
     2221 gagactttcc atttgaagat tacagtcaac atatatatag aaccatttct gatgggacta
     2281 tcagtgcatc caaacaggag agtgggaaga tggatgccaa aaccacagta ggaaaggcca
     2341 agggcaaatt gtggctcttt ggaaagaagc caaagggccc agcactagaa gatttcagtc
     2401 atctgccacc agaacagaga cgtaaaaaac tacagcagcg cattgatgaa cttaacagag
     2461 aactacagaa agaatcagac caaaaagatg cactcaacaa aatgaaagat gtatatgaga
     2521 agaatccaca aatgggggat ccagggagtt tgcagcctaa attagcagag accatgaata
     2581 acattgaccg cctacgaatg gaaatccata agaatgaggc ttggctctct gaagtcgaag
     2641 gcaaaacagg tgggagagga gacagaagac atagcagtga cataaatcat cttgtaacac
     2701 agggacgaga aagtcctgag ggaagttaca ctgatgatgc aaaccaggaa gtccgtgggc
     2761 caccccagca gcatggtcac cacaatgagt ttgatgatga atttgaggat gatgatccct
     2821 tgcctgctat tggacactgc aaagctatct acccttttga tggacataat gaaggtactc
     2881 tagcaatgaa agaaggtgaa gttctctaca ttatagagga ggacaaaggt gacggatgga
     2941 caagagctcg gagacagaac ggtgaagaag gctacgttcc cacgtcatac atagatgtaa
     3001 ctctagagaa aaacagtaaa ggttcctgag gcgcgcctct agaactatag tgagtcgtat
     3061 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3121 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3181 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3241 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     3301 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca
     3421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3481 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3541 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3601 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4321 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4441 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4501 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4621 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4801 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5341 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5461 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     5521 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5581 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5641 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5701 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5761 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5821 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5881 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5941 tgcgccgcta cagggcgcgt
//
