LOCUS       sequence-225613-        7089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  studying focal adhesion proteins by superresolution microscopy 
            (PALM) with dual labelling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  TITLE     Radenovic Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7089)
  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            "Unpublished"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: Paxillin_PSCFP2_ mEOS2
FEATURES             Location/Qualifiers
     source          1..7089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2374..3087
                     /codon_start=1
                     /product="photoswitchable cyan-to-green fluorescent 
                     protein"
                     /label=PS-CFP2
                     /note="mammalian codon-optimized"
                     /translation="MSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVATLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKTRAEVKFEGDTLVSRIELTGTDFKEDGNILGNKMEYNYNATNVYIVADKARNGIKV
                     NFKVRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYLE
                     FVTAAAITHGMDELYK"
     CDS             3112..3792
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    3914..4035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3951..3970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4005..4024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4042..4497)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4179..4200)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4391..4410
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4524..4628
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4630..4987
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4630..4821
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4653..4673)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4838..4973
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5022..5816
                     /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             5022..5816
                     /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(5076..5095)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5686..5705
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6004..6023)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6048..6095
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6424..7012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6913..6932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcaccatgg acgacctcga tgccttactg
      661 gcagacctgg aatccaccac ctcgcatatc tctaagcgac cggtgtttct aacagaggaa
      721 acgccttact cctacccaac tggaaaccat acgtaccagg agattgctgt gccaccgccg
      781 gtgcccccgc cgccttccag tgaggcccta aatggcactg tgattgaccc cttagaccag
      841 tggcaaccca gcgtgtccag atacggccac cagcaacctc agtcccagtc tccgatatac
      901 agctctagtg ccaaaagctc cagtgcctct gttcctagag acgggctcag ctccccttct
      961 ccccgtgcca gtgaggagga acacgtgtac agcttcccaa acaagcagaa gtctgcagaa
     1021 ccatctccca caatgaccag cacctccctg ggcagcaacc tctcggaact ggacaggctt
     1081 ctcctggaac tgaatgctgt tcaacataat ccccccagtg gcttctcagc agatgaagtc
     1141 agcagaagcc catcactgcc caatgtgacc ggacctcact atgtcatccc agagagcagc
     1201 agctctgcag gagggaaggc tgcaccccct acaaaagaaa aaccaaagcg gaatggtggg
     1261 cgtgggattg aagatgtgcg tcccagtgtg gagagcctgc tggatgagct ggagagctct
     1321 gtgccaagcc cagtccctgc aatcactgtg agccaagggg aggtgagcag cccccagcgt
     1381 gtcaacgcca gtcagcagca gacccgtatc tctgcttctt cagctacacg agaactggat
     1441 gagctgatgg cgtccctctc tgactttaag ttcatggcac aggggaaagc cggcggcagc
     1501 agctccccac catccaccac cccaaagcct ggcagtcagc tggacaccat gctgggaagt
     1561 ctgcagtctg acctgaacaa actgggggta gcgacggtcg ccaagggtgt gtgcggagcc
     1621 tgcaagaagc cgattgctgg gcaggtagtt acagccatgg ggaagacctg gcaccctgag
     1681 cacttcgtct gcacccactg ccaggaggag attggatcac ggaacttctt tgagcgggat
     1741 gggcagccct actgtgagaa ggactatcac aacctcttct ctcctcgctg ctactactgc
     1801 aatgggccaa tccttgataa agtggtgacg gctttggaca ggacgtggca ccctgaacac
     1861 tttttctgtg cccagtgtgg agttttcttt ggacctgaag gatttcatga gaaggatggc
     1921 aaagcctatt gccgcaagga ctactttgac atgtttgctc ccaagtgtgg aggctgtgcc
     1981 cgggctatcc tggaaaatta catttctgcc ttaaacaccc tgtggcaccc tgagtgcttt
     2041 gtctgtcggg aatgtttcac accgttcatc aacggcagct tcttcgagca cgacgggcag
     2101 ccctactgcg aggtgcatta ccacgagcgc cgcggctcgc tgtgctctgg ctgccagaag
     2161 cccatcacgg gacgctgcat cactgccatg ggcaagaaat ttcaccccga acactttgtc
     2221 tgtgccttct gcctcaagca gctcaacaaa ggaaccttca aagaacagaa cgacaagccc
     2281 tactgtcaga actgctttct caaactcttc tgtctcgagc tcaagcttcg aattcgcgaa
     2341 ccgtcagatc cgctagcgct accggtcgcc accatgagca agggcgccga gctgttcacc
     2401 ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg
     2461 agcggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     2521 accggcaagc tgcctgtgcc ctggcccacc ctggtggcca ccctgagcta cggcgtgcag
     2581 tgcttctcac gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct
     2641 gagggctaca tccaggagcg caccatcttc ttcgaggatg acggcaacta caagacgcgc
     2701 gccgaggtga agttcgaggg cgataccctg gtgagtcgca tcgagctgac cggcactgat
     2761 ttcaaggagg atggcaacat cctgggcaat aagatggagt acaactacaa cgccaccaat
     2821 gtgtacatcg tggccgacaa ggccaggaat ggcatcaagg tgaacttcaa ggtccgccac
     2881 aacatcaagg atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc
     2941 gatggccctg tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag
     3001 gaccccaacg agaagcgcga tcacatgatc tacctcgagt tcgtgaccgc cgccgccatc
     3061 acccacggca tggatgagct gtacaagtcc ggggatccac cggtcgccac catgagtgcg
     3121 attaagccag acatgaagat caaactccgt atggaaggca acgtaaacgg gcaccacttt
     3181 gtgatcgacg gagatggtac aggcaagcct tttgagggaa aacagagtat ggatcttgaa
     3241 gtcaaagagg gcggacctct gccttttgcc tttgatatcc tgaccactgc attccattac
     3301 ggcaacaggg tattcgccaa atatccagac aacatacaag actattttaa gcagtcgttt
     3361 cctaaggggt attcgtggga acgaagcttg actttcgaag acgggggcat ttgcattgcc
     3421 agaaacgaca taacaatgga aggggacact ttctataata aagttcgatt ttatggtacc
     3481 aactttcccg ccaatggtcc agttatgcag aagaagacgc tgaaatggga gccctccact
     3541 gagaaaatgt atgtgcgtga tggagtgctg acgggtgata ttcatatggc tttgttgctt
     3601 gaaggaaatg cccattaccg atgtgacttc agaactactt acaaagctaa ggagaagggt
     3661 gtcaagttac caggctacca ctttgtggac cactgcattg agattttaag ccatgacaaa
     3721 gattacaaca aggttaagct gtatgagcat gctgttgctc attctggatt gcctgacaat
     3781 gccagacgat aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     3841 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3901 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3961 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4021 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4081 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4141 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4201 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4261 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4321 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4381 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4441 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4501 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4561 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4621 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4681 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4741 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4801 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4861 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4921 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4981 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5041 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5101 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5161 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5221 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5281 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5341 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5401 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5461 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5521 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5581 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5641 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5701 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5761 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5821 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5881 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5941 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6001 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6061 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6121 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6181 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6241 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6301 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6361 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6421 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6481 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6541 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6601 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6661 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6721 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6781 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6841 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6901 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6961 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7021 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7081 tgcgttatc
//
