LOCUS       sequence-339652-        7041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of GFP-Cortactin. Note- This plasmid contains 
            GFP21, which has a sequence similar to YFP, but emission/excitation 
            similar to GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7041)
  AUTHORS   Yamada KM
  TITLE     Yamada Lab Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7041)
  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: 
            "unpublished"
COMMENT     Sequence Label: pGFP Cortactin
FEATURES             Location/Qualifiers
     source          1..7041
                     /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"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      911..920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(917..919,920..922,923..1633)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(962..983)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1223..1242)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1570..1591
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3328..3337
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3341..3475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3378..3397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3432..3451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3544..3901
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3544..3735
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3567..3587)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3752..3887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3921..3938)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3921..3937)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3930..3952)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4150..4605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4237..4256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4447..4468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4622..4641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4741..4763
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4801..4819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4887..4991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4992..5060,5061..5852)
                     /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     complement(5210..5229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6023..6611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6512..6531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6765..6782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6899..6920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6935..6940,6941..6958,6959..6965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6973..6989
                     /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)."
     primer_bind     6978..7000
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6997..7013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6997..7013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
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 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattcagcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      961 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1021 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1081 cgtgccctgg cccaccctcg tgaccacctt cgcctacggc ctgcagtgct tcgcccgcta
     1141 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1201 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1261 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1321 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1381 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1441 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1501 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1561 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1621 cgagctgtac aagggaggcg gatccatgtg gaaagcttca gcaggccacg ctgtgtccat
     1681 cgcccaggat gacgcggggg ccgatgactg ggagaccgac cctgattttg tgaatgatgt
     1741 gagtgagaag gagcaaagat ggggtgccaa gacggtgcag ggctccgggc accaggagca
     1801 tatcaacata cacaagctga gggagaatgt ctttcaagag catcagaccc ttaaggagaa
     1861 ggaacttgaa acaggaccaa aagcttccca tggctatgga gggaaatttg gtgtggaaca
     1921 agaccgaatg gataagtcag ctgtcggcca cgaatatcag tcgaaacttt ccaagcactg
     1981 ctcgcaggtg gactcggtcc gtggcttcgg aggcaagttt ggtgtccaga tggacagagt
     2041 tgatcagtct gctgtaggct ttgaatacca ggggaagact gagaagcatg cctcccagaa
     2101 agactactcc agtggttttg gcggcaagta tggcgtgcag gccgaccgag tagacaagag
     2161 cgcggtgggc ttcgactacc agggcaagac ggagaagcac gagtcacaga gagattactc
     2221 caaaggtttc ggcggcaaat acggtatcga caaggacaaa gtggataaga gcgccgttgg
     2281 ctttgagtat caaggcaaaa cggagaagca cgagtcccag aaagactatg tgaaagggtt
     2341 tggaggaaaa tttggtgtgc agacagacag acaagacaaa tgtgcccttg gctgggatca
     2401 ccaggagaaa ttgcagctgc atgaatccca aaaagattat aagactggtt ttggaggcaa
     2461 attcggtgtt cagtcggaga ggcaggactc cgctgctgtg gggtttgatt acaaggagaa
     2521 gctggccaag cacgagtccc agcaagacta ctccaaagga ttcggcggga agtatggggt
     2581 gcagaaggat cggatggata agaatgcgtc aacctttgag gatgtcaccc aggtgtcctc
     2641 tgcctaccag aagacagtac ctgtcgaagc tgtgaccagc aaaacaagta acatcagagc
     2701 taactttgaa aacctcgcta aggagaaaga gcaggaggac aggcggaagg cggaggcgga
     2761 gagagcccag cggatggcca aggagcggca ggagcaggaa gaggccagga ggaagctgga
     2821 ggagcaagcc agagccaaaa cgcaaacgcc ccctgtgtcg cccgcacctc agccaaccga
     2881 ggagaggctg ccctcgagcc ccgtctatga ggatgcggct tccttcaagg cagagctgag
     2941 ctacagaggc cctgtgagtg ggacggagcc ggagcccgtg tacagcatgg aggccgctga
     3001 ctaccgagag gccagcagcc agcagggcct ggcctatgcc acagaggctg tctatgaaag
     3061 cgcagaggcc ccgggccact atcccgcaga ggacagcacc tatgatgagt acgagaacga
     3121 tctggggatc acagccgtcg ccctgtacga ctaccaggct gcgggcgatg atgagatctc
     3181 atttgaccct gatgacatca tcaccaacat cgagatgatt gacgacggct ggtggcgcgg
     3241 ggtgtgcaag ggccggtacg ggctcttccc agccaactat gtggagctgc ggcagtagtc
     3301 tagagtcgac ctgcagaagc tagcttggcc gccatggccc aacttgttta ttgcagctta
     3361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa
     3481 ttaattcggc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3541 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3841 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3901 aaagctgtta acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3961 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4021 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     4081 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4141 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4201 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4261 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4321 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4381 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4441 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     4501 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4561 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4621 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4681 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4741 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4801 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4861 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4921 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5221 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5281 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5341 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5401 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5521 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5581 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5641 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5761 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5821 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5881 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5941 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6001 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6901 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6961 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta
     7021 cgaattaatt cgagctcgcc c
//
