LOCUS       sequence-160084-        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   Galisteo ML, Chernoff J, Su YC, Skolnik EY, Schlessinger J
  TITLE     The adaptor protein Nck links receptor tyrosine kinases with the 
            serine-threonine kinase Pak1.
  JOURNAL   J Biol Chem. 1996 Aug 30. 271(35):20997-1000.
  PUBMED    8798379
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: "J Biol 
            Chem. 1996 Aug 30. 271(35):20997-1000."
COMMENT     Sequence Label: pCMV6M-Pak1 P13A
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..33
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(578..595)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2198..2217)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2645..2665)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2650..2979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2830..2965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3017..3036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3018..3036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3050..3066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3050..3066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3063..3085)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3074..3090
                     /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(3098..3104,3105..3122,3123..3128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3143..3164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3281..3298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3452..4040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3532..3551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4211..5002,5003..5071))
                     /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     4834..4853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5072..5176)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5203..5658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5290..5309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5500..5521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5784..5806
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5798..5815
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5799..5815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5976..6355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6356..6559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6510..6530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggaacaga aactcatctc tgaagaggat ctgtctagag gatccatgtc aaataacggc
       61 ctagacattc aagacaagcc ggcagcccct ccgatgagaa ataccagcac tatgattgga
      121 gccggcagca aagatgctgg aaccctaaac catggttcta aacctctgcc tccaaaccca
      181 gaggagaaga aaaagaagga ccgattttac cgatccattt tacctggaga taaaacaaat
      241 aaaaagaaag agaaagagcg gccagagatt tctctccctt cagattttga acacacaatt
      301 catgtcggtt ttgatgctgt cacaggggag tttacgggaa tgccagagca gtgggcccgc
      361 ttgcttcaga catcaaatat cactaagtcg gagcagaaga aaaacccgca ggctgttctg
      421 gatgtgttgg agttttacaa ctcgaagaag acatccaaca gccagaaata catgagcttt
      481 acagataagt cagctgagga ttacaattct tctaatgcct tgaatgtgaa ggctgtgtct
      541 gagactcctg cagtgccacc agtttcagaa gatgaggatg atgatgatga tgatgctacc
      601 ccaccaccag tgattgctcc acgcccagag cacacaaaat ctgtatacac acggtctgtg
      661 attgaaccac ttcctgtcac tccaactcgg gacgtggcta catctcccat ttcacctact
      721 gaaaataaca ccactccacc agatgctttg acccggaata ctgagaagca gaagaagaag
      781 cctaaaatgt ctgatgagga gatcttggag aaattacgaa gcatagtgag tgtgggcgat
      841 cctaagaaga aatatacacg gtttgagaag attggacaag gtgcttcagg caccgtgtac
      901 acagcaatgg atgtggccac aggacaggag gtggccatta agcagatgaa tcttcagcag
      961 cagcccaaga aagagctgat tattaatgag atcctggtca tgagggaaaa caagaaccca
     1021 aacattgtga attacttgga cagttacctc gtgggagatg agctgtgggt tgttatggaa
     1081 tacttggctg gaggctcctt gacagatatg gtgacagaaa cttgcatgga tgaaggccaa
     1141 attgcagctg tgtgccgtga gtgtctgcag gctctggagt tcttgcattc gaaccaggtc
     1201 attcacagag acatcaagag tgacaatatt ctgttgggaa tggatagctc tgtcaagcta
     1261 actgactttg gattctgtgc acagataacc ccagagcaga gcaaacggag caccatggta
     1321 ggaaccccat actggatggc accagaggtt gtgacacgaa aggcctatgg gcccaaggtt
     1381 gacatctggt ccctgggcat catggccatc gaaatgattg aaggggagcc tccatacctc
     1441 aatgaaaacc ctctgagagc cttgtacctc attgccacca atgggacccc agaacttcag
     1501 aacccagaga agctgtcagc tatcttccgg gactttctga accgctgtct cgagatggat
     1561 gtggagaaga gaggttcagc taaagagctg atacagcatc aattcctgaa gattgccaag
     1621 cccctctcca gcctcactcc actgattgct gcagctaagg aggcaacaaa gaacaatcac
     1681 taaaaccaca ctcaccccag cctcattgtg ccaagccttc tgtgagataa atgcacattt
     1741 cagaaattcc aactcctgat gccctcttct ccttgccttg cttctcccat ttcctgatct
     1801 agcactcctc aagactttga tccttggaaa ccgtgtgtcc agcattgaag agaactgcaa
     1861 ctgaatgact aatcagatga tggccatttc taaataagga atttcctccc aattcatgga
     1921 tatgagggtg gtttatgatt aagggtttat ataaataaat gtttctagtc ttccgggcct
     1981 tcgtggcctc gaggaattcc gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     2041 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     2101 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     2161 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     2221 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     2281 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     2341 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     2401 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     2461 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     2521 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     2581 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg
     2641 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2701 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2761 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2821 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2881 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2941 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa
     3001 aaccagaaag ttaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt
     3061 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3121 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3181 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3241 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4141 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4201 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4261 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4321 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4381 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4441 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4501 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4561 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4621 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4681 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4741 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4801 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4861 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4921 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4981 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5041 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5101 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5161 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     5221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5401 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5521 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5641 tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5701 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5761 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     5821 gctgatctat acattgaatc aatattggca attagccata ttagtcattg gttatatagc
     5881 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     5941 ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga ctagttatta
     6001 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6061 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     6241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6301 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     6361 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6481 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     6541 ggtctatata agcagagctc gtttagtgaa ccgtcagaat taattcagat ctgctagcac
     6601 gcgtatcgat aagcttgcat gcctgcaggt cgac
//
