LOCUS       sequence-316715-        5806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin/Focal Adhesions, Excitation: 516, Emission: 529.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5806)
  TITLE     Davidson Yellow fluorescent proteins
REFERENCE   2  (bases 1 to 5806)
  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: mCitrine-VASP-5
FEATURES             Location/Qualifiers
     source          1..5806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(630..632,633..635,636..1346)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(675..696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(936..955)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1283..1304
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2611..2732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2648..2667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2702..2721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2739..3194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2876..2897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3088..3107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3221..3325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3327..3684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3327..3518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3350..3370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3535..3670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3719..4513
                     /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             3719..4513
                     /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(3773..3792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4383..4402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4701..4720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4745..4792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5121..5709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5610..5629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      661 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      721 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      781 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
      841 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      901 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      961 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1021 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1081 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1141 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1201 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
     1261 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1321 ctctcggcat ggacgagctg tacaagtccg gactcagatc tatgagcgag acggtcatct
     1381 gttccagccg ggctactgtg atgctttatg atgacagcaa caagcgatgg ctccctgctg
     1441 gcactggtcc ccaggccttc agccgcgtcc agatatacca caaccccact gctaactcct
     1501 tccgagttgt tggccgcaag atgcagccgg atcagcaggt ggttatcaac tgtgccatca
     1561 ttcggggtgt caagtacaat caggccactc ccatcttcca tcagtggcga gatgcccgcc
     1621 aggtctgggg cctcaacttc ggcagcaagg aggacgccat acagtttgca acaggcatgg
     1681 ccaacgccct agaggccttg gaaggaggtg ggcctccccc agccccagca ccccctgcct
     1741 ggtctgccca gaatggtccc tccccagagg agctggaaca acagaaaagg cagccggagc
     1801 atatggagcg ccgggtctcc aatgcaggag gcccacctgc tcccccagct gggggccctc
     1861 ctccacctcc aggacctccc cctcctccag gtcccccccc acccccaggt ctgccctcct
     1921 caggggtatc tggggcaggt catggagcag gggcagcccc accccctgca cccccactcc
     1981 ctacaacaca gggccccaat agtgggggtt ccggggcccc aggcctggct gctgccattg
     2041 ctggagccaa actcaggaaa gtgagcaagc aggaggaggc ctctgggggg cccctggccc
     2101 ccaaagctga gaacagtcga agcactggtg gggggcttat ggaagagatg aacgccatgc
     2161 tggcccggag aagaaaagcc acacaggttg gggagaagcc ccccaaagac gagtcagcca
     2221 gtgaggagtc agaggcccga ctccctgccc agagtgaacc tgtgagaaga ccctgggaga
     2281 agaacagcac aaccttgcca aggatgaagt cgtcctcttc tgtgactacc tccgaggccc
     2341 acccctcgac gccctgctcc agtgatgact ccgacttgga gagggtgaag caggagcttc
     2401 tggaagaggt gcggaaggag ctacagaaaa tgaaagagga aatcatcgaa gtctttgtcc
     2461 aggagctgag gaagcggggt tctccttgag cggccgcgac tctagatcat aatcagccat
     2521 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2581 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2701 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     2761 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2821 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2881 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2941 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3001 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3061 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3121 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3181 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3241 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3301 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3601 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3661 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3721 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3781 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3841 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3901 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3961 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4021 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4081 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4141 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4201 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4261 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4321 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4381 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4441 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4501 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4561 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4621 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4681 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4741 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4801 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4861 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4921 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4981 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5041 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5101 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5161 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5221 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5281 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5341 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5401 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5461 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5521 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5581 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5641 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5701 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5761 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataacc
//
