LOCUS       sequence-260200-        7881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7881)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 7881)
  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: mApple-Alpha-5-Integrin-12
FEATURES             Location/Qualifiers
     source          1..7881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        349..552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     503..523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3815..3824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3821..4531
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4234..4253
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4653..4774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4690..4709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4744..4763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4781..5236)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4918..4939)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5130..5149
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5263..5367
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5369..5726
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5369..5560
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5392..5412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5577..5712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5761..6555
                     /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             5761..6555
                     /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(5815..5834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6425..6444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6743..6762)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6787..6834
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7163..7751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7652..7671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
       61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      121 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      181 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      241 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      301 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      541 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg
      601 agctcaagct tcgaattctg cagtcgacgg taccaccatg gggagccgga cgccagagtc
      661 ccctctccac gccgtgcagc tgcgctgggg cccccggcgc cgacccccgc tgctgccgct
      721 gctgttgctg ctgctgccgc cgccacccag ggtcgggggc ttcaacttag acgcggaggc
      781 cccagcagta ctctcggggc ccccgggctc cttcttcgga ttctcagtgg agttttaccg
      841 gccgggaaca gacggggtca gtgtgctggt gggagcaccc aaggctaata ccagccagcc
      901 aggagtgctg cagggtggtg ctgtctacct ctgtccttgg ggtgccagcc ccacacagtg
      961 cacccccatt gaatttgaca gcaaaggctc tcggctcctg gagtcctcac tgtccagctc
     1021 agagggagag gagcctgtgg agtacaagtc cttgcagtgg ttcggggcaa cagttcgagc
     1081 ccatggctcc tccatcttgg catgcgctcc actgtacagc tggcgcacag agaaggagcc
     1141 actgagcgac cccgtgggca cctgctacct ctccacagat aacttcaccc gaattctgga
     1201 gtatgcaccc tgccgctcag atttcagctg ggcagcagga cagggttact gccaaggagg
     1261 cttcagtgcc gagttcacca agactggccg tgtggtttta ggtggaccag gaagctattt
     1321 ctggcaaggc cagatcctgt ctgccactca ggagcagatt gcagaatctt attaccccga
     1381 gtacctgatc aacctggttc aggggcagct gcagactcgc caggccagtt ccatctatga
     1441 tgacagctac ctaggatact ctgtggctgt tggtgaattc agtggtgatg acacagaaga
     1501 ctttgttgct ggtgtgccca aagggaacct cacttacggc tatgtcacca tccttaatgg
     1561 ctcagacatt cgatccctct acaacttctc aggggaacag atggcctcct actttggcta
     1621 tgcagtggcc gccacagacg tcaatgggga cgggctggat gacttgctgg tgggggcacc
     1681 cctgctcatg gatcggaccc ctgacgggcg gcctcaggag gtgggcaggg tctacgtcta
     1741 cctgcagcac ccagccggca tagagcccac gcccaccctt accctcactg gccatgatga
     1801 gtttggccga tttggcagct ccttgacccc cctgggggac ctggaccagg atggctacaa
     1861 tgatgtggcc atcggggctc cctttggtgg ggagacccag cagggagtag tgtttgtatt
     1921 tcctgggggc ccaggagggc tgggctctaa gccttcccag gttctgcagc ccctgtgggc
     1981 agccagccac accccagact tctttggctc tgcccttcga ggaggccgag acctggatgg
     2041 caatggatat cctgatctga ttgtggggtc ctttggtgtg gacaaggctg tggtatacag
     2101 gggccgcccc atcgtgtccg ctagtgcctc cctcaccatc ttccccgcca tgttcaaccc
     2161 agaggagcgg agctgcagct tagaggggaa ccctgtggcc tgcatcaacc ttagcttctg
     2221 cctcaatgct tctggaaaac acgttgctga ctccattggt ttcacagtgg aacttcagct
     2281 ggactggcag aagcagaagg gaggggtacg gcgggcactg ttcctggcct ccaggcaggc
     2341 aaccctgacc cagaccctgc tcatccagaa tggggctcga gaggattgca gagagatgaa
     2401 gatctacctc aggaacgagt cagaatttcg agacaaactc tcgccgattc acatcgctct
     2461 caacttctcc ttggaccccc aagccccagt ggacagccac ggcctcaggc cagccctaca
     2521 ttatcagagc aagagccgga tagaggacaa ggctcagatc ttgctggact gtggagaaga
     2581 caacatctgt gtgcctgacc tgcagctgga agtgtttggg gagcagaacc atgtgtacct
     2641 gggtgacaag aatgccctga acctcacttt ccatgcccag aatgtgggtg agggtggcgc
     2701 ctatgaggct gagcttcggg tcaccgcccc tccagaggct gagtactcag gactcgtcag
     2761 acacccaggg aacttctcca gcctgagctg tgactacttt gccgtgaacc agagccgcct
     2821 gctggtgtgt gacctgggca accccatgaa ggcaggagcc agtctgtggg gtggccttcg
     2881 gtttacagtc cctcatctcc gggacactaa gaaaaccatc cagtttgact tccagatcct
     2941 cagcaagaat ctcaacaact cgcaaagcga cgtggtttcc tttcggctct ccgtggaggc
     3001 tcaggcccag gtcaccctga acggtgtctc caagcctgag gcagtgctat tcccagtaag
     3061 cgactggcat ccccgagacc agcctcagaa ggaggaggac ctgggacctg ctgtccacca
     3121 tgtctatgag ctcatcaacc aaggccccag ctccattagc cagggtgtgc tggaactcag
     3181 ctgtccccag gctctggaag gtcagcagct cctatatgtg accagagtta cgggactcaa
     3241 ctgcaccacc aatcacccca ttaacccaaa gggcctggag ttggatcccg agggttccct
     3301 gcaccaccag caaaaacggg aagctccaag ccgcagctct gcttcctcgg gacctcagat
     3361 cctgaaatgc ccggaggctg agtgtttcag gctgcgctgt gagctcgggc ccctgcacca
     3421 acaagagagc caaagtctgc agttgcattt ccgagtctgg gccaagactt tcttgcagcg
     3481 ggagcaccag ccatttagcc tgcagtgtga ggctgtgtac aaagccctga agatgcccta
     3541 ccgaatcctg cctcggcagc tgccccaaaa agagcgtcag gtggccacag ctgtgcaatg
     3601 gaccaaggca gaaggcagct atggcgtccc actgtggatc atcatcctag ccatcctgtt
     3661 tggcctcctg ctcctaggtc tactcatcta catcctctac aagcttggat tcttcaaacg
     3721 ctccctccca tatggcaccg ccatggaaaa agctcagctc aagcctccag ccacctctga
     3781 tgccacggta ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga
     3841 gaataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     3901 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca
     3961 gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     4021 ccctcagttc atgtacggct ccaaggtcta cattaagcac ccagccgaca tccccgacta
     4081 cttcaagctg tccttccccg agggcttcag gtgggagcgc gtgatgaact tcgaggacgg
     4141 cggcattatt cacgttaacc aggactcctc cctgcaggac ggcgtgttca tctacaaggt
     4201 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     4261 ctgggaggcc tccgaggagc ggatgtaccc cgaggacggc gccctgaaga gcgagatcaa
     4321 gaagaggctg aagctgaagg acggcggcca ctacgccgcc gaggtcaaga ccacctacaa
     4381 ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc gacatcaagt tggacatcgt
     4441 gtcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     4501 caccggcggc atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc
     4561 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4621 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4681 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4741 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4801 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4861 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4921 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4981 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5041 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5101 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5161 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5221 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5281 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5341 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     5401 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5461 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5521 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5581 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5641 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5701 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5761 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5821 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5881 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5941 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6001 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6061 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6121 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6181 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6241 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     6301 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6361 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6421 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6481 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6541 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6601 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6661 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6721 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6781 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6841 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6901 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6961 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     7021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     7861 tgcattagtt attaatagta a
//
