LOCUS       Exported                4696 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Localization: N1 Cloning Vector, Excitation: 505 / 569, Emission: 
            516 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    mEos4b-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4696)
  AUTHORS   Paez-Segala MG, Sun MG, Shtengel G, Viswanathan S, Baird MA, Macklin
            JJ, Patel R, Allen JR, Howe ES, Piszczek G, Hess HF, Davidson MW, 
            Wang Y, Looger LL
  TITLE     Fixation-resistant photoactivatable fluorescent proteins for CLEM.
  JOURNAL   Nat Methods. 2015 Jan 12. doi: 10.1038/nmeth.3225.
  PUBMED    25581799
REFERENCE   2  (bases 1 to 4696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4696
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     misc_feature    598..678
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      680..689
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             686..1369
                     /codon_start=1
                     /product="fixation-resistant truly monomeric green-to-red 
                     photoswitchable fluorescent protein (Paez-Segala et al., 
                     2015)"
                     /label=mEos4b
                     /note="derived from mEos2"
                     /translation="MVSAIKPDMRIKLRMEGNVNGHHFVIDGDGTGKPYEGKQTMDLEV
                     KEGGPLPFAFDILTTAFHYGNRVFVKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNA
                     RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL
                     LEGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     polyA_signal    1491..1612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1528..1547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1582..1601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1619..2074)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1756..1777)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1968..1987
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2101..2205
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2207..2564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2230..2250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2415..2550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2477..2496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2599..3393
                     /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(2653..2672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3263..3282
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3581..3600)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3625..3672
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4001..4589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4490..4509
                     /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 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
      661 ggcccgggat ccaccggtcg ccaccatggt gagtgcgatt aagccagaca tgaggatcaa
      721 actccgtatg gaaggcaacg taaacgggca ccactttgtg atcgacggag atggtacagg
      781 caagccttat gagggaaaac agaccatgga tcttgaagtc aaagagggcg gacctctgcc
      841 ttttgccttt gatatcctga ccactgcatt ccattacggc aacagggtat tcgtgaaata
      901 tccagacaac atacaagact attttaagca gtcgtttcct aaggggtatt cgtgggaacg
      961 aagcttgact ttcgaagacg ggggcatttg caatgccaga aacgacataa caatggaagg
     1021 ggacactttc tataataaag ttcgatttta tggtaccaac tttcccgcca atggtccagt
     1081 tatgcagaag aagacgctga aatgggagcc ctccactgag aaaatgtatg tgcgtgatgg
     1141 agtgctgacg ggtgatattg agatggcttt gttgcttgaa ggaaatgccc attaccgatg
     1201 tgacttcaga actacttaca aagctaagga gaagggtgtc aagttaccag gcgcccactt
     1261 tgtggaccac gccattgaga ttttaagcca tgacaaagat tacaacaagg ttaagctgta
     1321 tgagcatgct gttgctcatt ctggattgcc tgacaatgcc agacgataag cggccgcgac
     1381 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1441 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     1621 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1681 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1741 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1801 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     1861 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     1921 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     1981 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2041 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2101 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2161 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2221 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2281 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2581 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2641 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2701 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2761 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2821 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2881 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2941 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3001 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3061 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3121 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3181 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3241 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3301 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3361 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3421 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3481 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3541 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3601 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3661 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3721 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3781 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     3841 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3901 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3961 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4021 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4081 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4141 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4201 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4261 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4321 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4381 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4441 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4501 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4561 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4621 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4681 ataaccgtat taccgc
//
