LOCUS       sequence-203086-        7075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 7075)
  AUTHORS   Yamada KM
  TITLE     Yamada Lab Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7075)
  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            "unpublished"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: pRK GFP Paxillin
FEATURES             Location/Qualifiers
     source          1..7075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1020..1399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1400..1603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1554..1574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1600..1624
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1834..1852
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1834..1851
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1865..1884
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1930..1939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1936..1938,1939..1941,1942..2652)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1981..2002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2242..2261)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2589..2610
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      4381..4390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4394..4528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4431..4450)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4485..4504
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4597..4954
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4597..4788
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4620..4640)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4805..4940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4974..4991)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4974..4990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..5005)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     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     complement(5675..5694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5794..5816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5854..5872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5940..6044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6045..6113,6114..6905)
                     /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(6263..6282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg
     1021 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     1441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     1501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     1561 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     1621 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     1681 cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt
     1741 aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa
     1801 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaataacatc
     1861 cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct cggttctatc
     1921 gattcagccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1981 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2041 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2101 gtgccctggc ccaccctcgt gaccaccttc gcctacggcc tgcagtgctt cgcccgctac
     2161 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2221 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2281 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2341 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2401 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2461 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2521 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2581 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2641 gagctgtaca agggaggcgg atccaagctt atggacgacc tcgacgccct gctggcggac
     2701 ttggagtcta ccacctccca catctccaaa cggcctgtgt tcttgtcgga ggagaccccc
     2761 tactcatacc caactggaaa ccacacatac caggagattg ccgtgccacc ccccgtcccc
     2821 ccacccccgt ccagcgaggc cctcaatggc acaatccttg accccttaga ccagtggcag
     2881 cccagcggct cccgattcat ccaccagcag cctcagtcct catcacctgt gtacggctcc
     2941 agtgccaaaa cttccagtgt ctccaaccct caggacagtg ttggctctcc gtgctcccga
     3001 gtgggtgagg aggagcacgt ctacagcttc cccaacaagc agaaatcagc tgagccttca
     3061 cccaccgtaa tgagcacgtc cctgggcagc aacctttctg aactcgaccg cctgctgctg
     3121 gaactgaacg ctgtacagca taacccgcca ggcttccctg cagatgaggc caactcaagc
     3181 cccccgcttc ctggggccct gagccccctc tatggtgtcc cagagactaa cagccccttg
     3241 ggaggcaaag ctgggcccct gacgaaagag aagcctaagc ggaatggggg ccggggcctg
     3301 gaggacgtgc ggcccagtgt ggagagtctc ttggatgaac tggagagctc cgtgcccagc
     3361 cccgtccctg ccatcactgt gaaccagggc gagatgagca gcccgcagcg cgtcacctcc
     3421 acccaacagc agacacgcat ctcggcctcc tctgccacca gggagctgga cgagctgatg
     3481 gcttcgctgt cggatttcaa gttcatggcc caggggaaga cagggagcag ctcaccccct
     3541 ggggggcccc cgaagcccgg gagccagctg gacagcatgc tggggagcct gcagtctgac
     3601 ctgaacaagc tgggggtcgc cacagtcgcc aaaggagtct gcggggcctg caagaagccc
     3661 atcgccgggc aggttgtgac cgccatgggg aagacgtggc accccgagca cttcgtctgc
     3721 acccactgcc aggaggagat cggatcccgg aacttcttcg agcgggatgg acagccctac
     3781 tgtgaaaagg actaccacaa cctcttctcc ccgcgctgct actactgcaa cggccccatc
     3841 ctggataaag tggtgacagc ccttgaccgg acgtggcacc ctgaacactt cttctgtgca
     3901 cagtgtggag ccttctttgg tcccgaaggg ttccacgaga aggacggcaa ggcctactgt
     3961 cgcaaggact acttcgacat gttcgcaccc aagtgtggcg gctgcgcccg ggccatcctg
     4021 gagaactata tctcagccct caacacgctg tggcatcctg agtgctttgt gtgccgggaa
     4081 tgcttcacgc cattcgtgaa cggcagcttc ttcgagcacg acgggcagcc ctactgtgag
     4141 gtgcactacc acgagcggcg cggctcgctg tgttctggct gccagaagcc catcaccggc
     4201 cgctgcatca ccgccatggc caagaagttc caccccgagc acttcgtctg tgccttctgc
     4261 ctcaagcagc tcaacaaggg caccttcaag gagcagaacg acaagcctta ctgtcagaac
     4321 tgcttcctca agctcttctg ctaggctcga gtctagagtc gacctgcaga agctagcttg
     4381 gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4441 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4501 catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac catggcctga
     4561 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     4621 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4681 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4801 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4861 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4921 aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt ggcactggcc
     4981 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     5041 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5101 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     5161 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     5221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5401 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5521 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     5581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5641 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5701 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5761 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5821 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5881 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5941 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6001 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6061 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6121 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6181 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6241 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6301 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6361 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6421 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6481 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6541 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6601 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6661 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6721 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6781 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6841 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6901 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6961 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7021 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
