LOCUS       sequence-138985-        4701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 490 / 553, Emission: 507 / 573.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4701)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4701)
  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            "unpublished"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: Dendra2-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..4701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             610..660
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             682..1374
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     polyA_signal    1496..1617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1533..1552)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1587..1606
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1624..2079)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1761..1782)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1973..1992
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2106..2210
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2212..2569
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2212..2403
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2235..2255)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2420..2555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2604..3398
                     /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             2604..3398
                     /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(2658..2677)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3268..3287
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3586..3605)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3630..3677
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4006..4594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4495..4514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgccacca tgggcgtggc cgacttgatc aagaagttcg agtccatctc caaggaggag
      661 ggggatccac cggtcgccac catgaacacc ccgggaatta acctgatcaa ggaggacatg
      721 cgcgtgaagg tgcacatgga gggcaacgtg aacggccacg ccttcgtgat cgagggcgag
      781 ggcaagggca agccctacga gggcacccag accgccaacc tgaccgtgaa ggagggcgcc
      841 cccctgccct tcagctacga catcctgacc accgccgtgc actacggcaa ccgggtgttc
      901 accaagtacc ccgaggacat ccccgactac ttcaagcaga gcttccccga gggctacagc
      961 tgggagcgca ccatgacctt cgaggacaag ggcatctgca ccatccgcag cgacatcagc
     1021 ctggagggcg actgcttctt ccagaacgtg cgcttcaagg gcaccaactt cccccccaac
     1081 ggccccgtga tgcagaagaa gaccctgaag tgggagccca gcaccgagaa gctgcacgtg
     1141 cgcgacggcc tgctggtggg caacatcaac atggccctgc tgctggaggg cggcggccac
     1201 tacctgtgcg acttcaagac cacctacaag gccaagaagg tggtgcagct gcccgacgcc
     1261 cacttcgtgg accaccgcat cgagatcctg ggcaacgaca gcgactacaa caaggtgaag
     1321 ctgtacgagc acgccgtggc ccgctacagc cccctgccca gccaggtgtg gtaagcggcc
     1381 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1441 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1501 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1561 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     1621 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1681 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1741 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1801 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1861 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1921 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1981 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2041 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2101 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2161 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2221 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2281 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2341 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2401 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2461 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2521 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     2581 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2641 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2701 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2761 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     2821 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2881 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2941 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3001 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3061 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3121 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3181 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3241 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3301 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3361 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3421 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3481 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3541 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     3601 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3661 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3721 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3781 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3841 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3901 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3961 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4021 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4081 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4141 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4201 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4261 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4321 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4381 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4441 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4501 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4561 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4621 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4681 tgtggataac cgtattaccg c
//
