LOCUS       sequence-226620-        6396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 406, Emission: 526.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6396)
  TITLE     Davidson Yellow fluorescent proteins
REFERENCE   2  (bases 1 to 6396)
  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"
COMMENT     Sequence Label: mAmetrine-Paxillin-22
FEATURES             Location/Qualifiers
     source          1..6396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2217..2269
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2277..2286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2283..2285,2286..2288,2289..3002)
                     /codon_start=1
                     /product="yellow fluorescent protein with a large Stokes 
                     shift, excitable with violet light (Ai et al., 2008)"
                     /label=mAmetrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTSGKLPVPWPTLVTTLSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GSYRTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNMNVWDAYITADKQKNGIK
                     ANFKIEHNVEDGGVQLADAYQQNTPIGDGSVLLPDNHYLSFQSKLFKDPNEQRDHMVLL
                     EFVTAAGITPGMDELYK"
     primer_bind     complement(2328..2349)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3124..3245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3161..3180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3215..3234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3252..3707)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3389..3410)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3601..3620
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3734..3838
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3840..4197
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3840..4031
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3863..3883)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4048..4183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4232..5026
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4232..5026
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4286..4305)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4896..4915
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5214..5233)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5258..5305
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5634..6222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6123..6142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggacgacct cgatgcctta ctggcagacc tggaatccac cacctcgcat atctctaagc
      601 gaccggtgtt tctaacagag gaaacgcctt actcctaccc aactggaaac catacgtacc
      661 aggagattgc tgtgccaccg ccggtgcccc cgccgccttc cagtgaggcc ctaaatggca
      721 ctgtgattga ccccttagac cagtggcaac ccagcgtgtc cagatacggc caccagcaac
      781 ctcagtccca gtctccgata tacagctcta gtgccaaaag ctccagtgcc tctgttccta
      841 gagacgggct cagctcccct tctccccgtg ccagtgagga ggaacacgtg tacagcttcc
      901 caaacaagca gaagtctgca gaaccatctc ccacaatgac cagcacctcc ctgggcagca
      961 acctctcgga actggacagg cttctcctgg aactgaatgc tgttcaacat aatcccccca
     1021 gtggcttctc agcagatgaa gtcagcagaa gcccatcact gcccaatgtg accggacctc
     1081 actatgtcat cccagagagc agcagctctg caggagggaa ggctgcaccc cctacaaaag
     1141 aaaaaccaaa gcggaatggt gggcgtggga ttgaagatgt gcgtcccagt gtggagagcc
     1201 tgctggatga gctggagagc tctgtgccaa gcccagtccc tgcaatcact gtgagccaag
     1261 gggaggtgag cagcccccag cgtgtcaacg ccagtcagca gcagacccgt atctctgctt
     1321 cttcagctac acgagaactg gatgagctga tggcgtccct ctctgacttt aagttcatgg
     1381 cacaggggaa agccggcggc agcagctccc caccatccac caccccaaag cctggcagtc
     1441 agctggacac catgctggga agtctgcagt ctgacctgaa caaactgggg gtagcgacgg
     1501 tcgccaaggg tgtgtgcgga gcctgcaaga agccgattgc tgggcaggta gttacagcca
     1561 tggggaagac ctggcaccct gagcacttcg tctgcaccca ctgccaggag gagattggat
     1621 cacggaactt ctttgagcgg gatgggcagc cctactgtga gaaggactat cacaacctct
     1681 tctctcctcg ctgctactac tgcaatgggc caatccttga taaagtggtg acggctttgg
     1741 acaggacgtg gcaccctgaa cactttttct gtgcccagtg tggagttttc tttggacctg
     1801 aaggatttca tgagaaggat ggcaaagcct attgccgcaa ggactacttt gacatgtttg
     1861 ctcccaagtg tggaggctgt gcccgggcta tcctggaaaa ttacatttct gccttaaaca
     1921 ccctgtggca ccctgagtgc tttgtctgtc gggaatgttt cacaccgttc atcaacggca
     1981 gcttcttcga gcacgacggg cagccctact gcgaggtgca ttaccacgag cgccgcggct
     2041 cgctgtgctc tggctgccag aagcccatca cgggacgctg catcactgcc atgggcaaga
     2101 aatttcaccc cgaacacttt gtctgtgcct tctgcctcaa gcagctcaac aaaggaacct
     2161 tcaaagaaca gaacgacaag ccctactgtc agaactgctt tctcaaactc ttctgtctcg
     2221 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     2281 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2341 acggcgacgt aaacggccac aagttcagcg tgcgcggcga gggcgagggc gatgccacca
     2401 acggcaagct gaccctgaag ttcatctgca cctccggcaa gctgcccgtg ccctggccca
     2461 ccctcgtgac caccctgtct tacggcgtgc agtgcttcgc ccgctacccc gaccacatga
     2521 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2581 ccttcaagga cgacggcagc tacaggaccc gcgccgaggt gaagttcgag ggcgacaccc
     2641 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2701 acaagctgga gtacaacatg aacgtgtggg acgcgtatat cacggccgac aagcagaaga
     2761 acggcatcaa agcgaacttc aagatcgagc acaacgtcga ggacggcggc gtgcagctcg
     2821 ccgacgcgta ccagcagaac acccccatcg gcgacggctc cgtgctgctg cctgacaacc
     2881 actacctgag cttccagagc aagctgttca aagaccccaa cgagcagcgc gatcacatgg
     2941 tcctgctgga gttcgttacc gccgccggga tcactcccgg catggacgag ctgtacaagt
     3001 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     3061 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3121 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3181 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3241 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3301 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3361 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3421 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3481 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3541 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3601 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3661 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3721 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3781 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3841 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4021 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4081 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4141 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4201 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4261 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4321 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4381 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4441 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4501 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4561 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4621 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4681 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4741 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4801 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4861 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4921 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4981 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5041 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5101 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5161 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5221 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5281 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5341 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5401 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5461 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5761 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6301 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     6361 ggggtcatta gttcatagcc catatatgga gttccg
//
