LOCUS       sequence-203261-        4692 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: C1 Cloning Vector, Excitation: 507 / 572, Emission: 
            516 / 580.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  AUTHORS   Zhang M, Chang H, Zhang Y, Yu J, Wu L, Ji W, Chen J, Liu B, Lu J, 
            Liu Y, Zhang J, Xu P, Xu T
  TITLE     Rational design of true monomeric and bright photoactivatable 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2012 May 13;9(7):727-9. doi: 10.1038/nmeth.2021.
  PUBMED    22581370
REFERENCE   2  (bases 1 to 4692)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-05-13- 13"; volume: "9"; 
            issue: "7"; pages: "727-9"
COMMENT     Sequence Label: mEos3.2-C1
FEATURES             Location/Qualifiers
     source          1..4692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             568..1245
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     misc_feature    1246..1311
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1435..1556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1472..1491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1526..1545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1563..2018)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1700..1721)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1912..1931
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2045..2149
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2151..2508
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2151..2342
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2174..2194)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2359..2494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2543..3337
                     /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             2543..3337
                     /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(2597..2616)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3207..3226
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3525..3544)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3569..3616
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3945..4533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4434..4453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccggt cgccaccatg agtgcgatta agccagacat gaagatcaaa
      601 ctccgtatgg aaggcaacgt aaacgggcac cactttgtga tcgacggaga tggtacaggc
      661 aagccttttg agggaaaaca gagtatggat cttgaagtca aagagggcgg acctctgcct
      721 tttgcctttg atatcctgac cactgcattc cattacggca acagggtatt cgccaaatat
      781 ccagacaaca tacaagacta ttttaagcag tcgtttccta aggggtattc gtgggaacga
      841 agcttgactt tcgaagacgg gggcatttgc aacgccagaa acgacataac aatggaaggg
      901 gacactttct ataataaagt tcgattttat ggtaccaact ttcccgccaa tggtccagtt
      961 atgcagaaga agacgctgaa atgggagccc tccactgaga aaatgtatgt gcgtgatgga
     1021 gtgctgacgg gtgatattga gatggctttg ttgcttgaag gaaatgccca ttaccgatgt
     1081 gacttcagaa ctacttacaa agctaaggag aagggtgtca agttaccagg cgcccacttt
     1141 gtggaccact gcattgagat tttaagccat gacaaagatt acaacaaggt taagctgtat
     1201 gagcatgctg ttgctcattc tggattgcct gacaatgcca gacgatccgg actcagatct
     1261 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct
     1321 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1381 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     1561 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1621 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     1681 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1741 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1801 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1861 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1921 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1981 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2041 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2101 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2161 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2341 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2401 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2461 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     2521 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2581 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2641 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2701 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     2761 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2821 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2881 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2941 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3001 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3061 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3121 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3181 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3241 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3301 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3361 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3421 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3481 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     3541 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     3601 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3661 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     3721 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     3781 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3841 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3901 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3961 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4021 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4081 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4141 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4201 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4261 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4321 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4381 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4441 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4501 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4561 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4621 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     4681 agttcatagc cc
//
