LOCUS       sequence-204937-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Lyn-mTurquoise2-VASP at the plasma membrane of mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  AUTHORS   O'Banion CP, Priestman MA, Hughes RM, Herring LE, Capuzzi SJ, 
            Lawrence DS
  TITLE     Design and Profiling of a Subcellular Targeted Optogenetic 
            cAMP-Dependent Protein Kinase.
  JOURNAL   Cell Chem Biol. 2017 Oct 25. pii: S2451-9456(17)30355-0. doi: 
            10.1016/j.chembiol.2017.09.011.
  PUBMED    29104065
REFERENCE   2  (bases 1 to 5563)
  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: "Cell Chem 
            Biol. 2017 Oct 25. pii: S2451-9456(17)30355-0. doi: 
            10.1016/j.chembiol.2017.09.011."
COMMENT     Sequence Label: PM PKA Reporter
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..1346
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          395..1337
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1294..1314
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1392..1401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1434..1460
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1461..1463,1464..1466,1467..2177)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1506..1527)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1767..1786)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2114..2135
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2351..2472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2388..2407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2442..2461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2479..2934)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2616..2637)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2828..2847
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2961..3065
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3067..3424
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3067..3258
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3090..3110)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3275..3410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3459..4253
                     /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             3459..4253
                     /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(3513..3532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4123..4142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4441..4460)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4485..4532
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4861..5449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5350..5369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tgagtaattc atacaaaagg actcgcccct gccttgggga atcccaggga
       61 ccgtcgttaa actcccacta acgtagaacc cagagatcgc tgcgttcccg ccccctcacc
      121 cgcccgctct cgtcatcact gaggtggaga agagcatgcg tgaggctccg gtgcccgtca
      181 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      241 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      301 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      361 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      421 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cgcccctggc
      481 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      541 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggctt gggcgctggg
      601 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      661 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      721 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      781 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
      841 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
      901 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
      961 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     1021 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1081 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1141 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1201 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1261 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1321 tttttcttcc atttcaggtg tcgtgacgct agcgctaccg gactcagatc tcgagctcaa
     1381 gcttcgaatt cgccaccatg ggctgcatca agagcaaggg caaggacagc gcctacccat
     1441 acgatgttcc agattacgct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1501 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1561 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1621 tgcccgtgcc ctggcccacc ctcgtgacca ccctgtcctg gggcgtgcag tgcttcgccc
     1681 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1741 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1801 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1861 acggcaacat cctggggcac aagctggagt acaactactt tagcgacaac gtctatatca
     1921 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1981 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2041 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     2101 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2161 tggacgagct gtacaagagc gagcacatcg agcggcgggt gagcaacgcc ggcggccccc
     2221 ccgcctgagg atcccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2281 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2341 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2401 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2461 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2521 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2581 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2641 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2701 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2761 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2821 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2881 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2941 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3001 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3061 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3181 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3241 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3301 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3361 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3421 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3961 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4261 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4441 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4501 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4561 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4621 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4681 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5521 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
