LOCUS       sequence-365722-        7327 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Localization: Kinetochore, Excitation: 505 / 569, Emission: 516 / 
            581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7327)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 7327)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: tdEos-Hec1-C-12
FEATURES             Location/Qualifiers
     source          1..7327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2530..3207
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             3256..3933
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     polyA_signal    4055..4176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4092..4111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4146..4165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4183..4638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4320..4341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4532..4551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4665..4769
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4771..5128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4771..4962
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4794..4814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4979..5114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5163..5957
                     /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             5163..5957
                     /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(5217..5236)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5827..5846
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6145..6164)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6189..6236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6565..7153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7054..7073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct cgccaccatg aagcgcagtt cagtttccag cggtggtgct
      601 ggccgcctct ccatgcagga gttaagatcc caggatgtaa ataaacaagg cctctatacc
      661 cctcaaacca aagagaaacc aacctttgga aagttgagta taaacaaacc gacatctgaa
      721 agaaaagtct cgctatttgg caaaagaact agtggacatg gatcacggaa tagtcaactt
      781 ggtatatttt ccagttctga gaaaatcaag gacccgagac cacttaacga caaagcattc
      841 attcagcagt gtattcgaca actctgtgag tttcttacag aaaatggtta tgcacataat
      901 gtgtccatga aatctctaca agctccctct gttaaagact tcctgaagat cttcacattt
      961 ctttatggct tcctgtgccc ctcatacgaa cttcctgaca caaagtttga agaagaggtt
     1021 ccaagaatct ttaaagacct tgggtatcct tttgcactat ccaaaagctc catgtacaca
     1081 gtgggggctc ctcatacatg gcctcacatt gtggcagcct tagtttggct aatagactgc
     1141 atcaagatac atactgccat gaaagaaagc tcacctttat ttgatgatgg gcagccttgg
     1201 ggagaagaaa ctgaagatgg aattatgcat aataagttgt ttttggacta caccataaaa
     1261 tgctatgaga gttttatgag tggtgccgac agctttgatg agatgaatgc agagctgcag
     1321 tcaaaactga aggatttatt taatgtggat gcttttaagc tggaatcatt agaagcaaaa
     1381 aacagagcat tgaatgaaca gattgcaaga ttggaacaag aaagagaaaa agaaccgaat
     1441 cgtctagagt cgttgagaaa actgaaggct tccttacaag gagatgttca aaagtatcag
     1501 gcatacatga gcaatttgga gtctcattca gccattcttg accagaaatt aaatggtctc
     1561 aatgaggaaa ttgctagagt agaactagaa tgtgaaacaa taaaacagga gaacactcga
     1621 ctacagaata tcattgacaa ccagaagtac tcagttgcag acattgagcg aataaatcat
     1681 gaaagaaatg aattgcagca gactattaat aaattaacca aggacctgga agctgaacaa
     1741 cagaagttgt ggaatgagga gttaaaatat gccagaggca aagaagcgat tgaaacacaa
     1801 ttagcagagt atcacaaatt ggctagaaaa ttaaaactta ttcctaaagg tgctgagaat
     1861 tccaaaggtt atgactttga aattaagttt aatcccgagg ctggtgccaa ctgccttgtc
     1921 aaatacaggg ctcaagttta tgtacctctt aaggaactcc tgaatgaaac tgaagaagaa
     1981 attaataaag ccctaaataa aaaaatgggt ttggaggata ctttagaaca attgaatgca
     2041 atgataacag aaagcaagag aagtgtgaga actctgaaag aagaagttca aaagctggat
     2101 gatctttacc aacaaaaaat taaggaagca gaggaagagg atgaaaaatg tgccagtgag
     2161 cttgagtcct tggagaaaca caagcacctg ctagaaagta ctgttaacca ggggctcagt
     2221 gaagctatga atgaattaga tgctgttcag cgggaatacc aactagttgt gcaaaccacg
     2281 actgaagaaa gacgaaaagt gggaaataac ttgcaacgtc tgttagagat ggttgctaca
     2341 catgttgggt ctgtagagaa acatcttgag gagcagattg ctaaagttga tagagaatat
     2401 gaagaatgca tgtcagaaga tctctcggaa aatattaaag agattagaga taagtatgag
     2461 aagaaagcta ctctaattaa gtcttctgaa gaagcggtac cgcgggcccg ggatccaccg
     2521 gtcgccacca tgagtgcgat taagccagac atgaagatca acctccgtat ggaaggcaac
     2581 gtaaacgggc accactttgt gatcgacgga gatggtacag gcaagccttt tgagggaaaa
     2641 cagagtatgg atcttgaagt caaagagggc ggacctctgc cttttgcctt tgatatcctg
     2701 accactgcat tccattacgg caacagggta ttcgccgaat atccagacca catacaagac
     2761 tattttaagc agtcgtttcc taaggggtat tcgtgggaac gaagcttgac tttcgaagac
     2821 gggggcattt gcattgccag aaacgacata acaatggaag gggacacttt ctataataaa
     2881 gttcgatttc acggtgtaaa ctttcccgcc aatggtccag ttatgcagaa gaagacgctg
     2941 aaatgggagc cctccactga gaaaatgtat gtgcgtgatg gagtgctgac tggtgatatt
     3001 cgcatggctt tgttgcttga aggaaatgcc cattaccgat gtgacttcag aactacttac
     3061 aaagctaagg agaagggtgt caagttacca ggctaccact ttgtggacca ctgcattgag
     3121 attttaagcc atgacaaaga ttacaacaag gttaagctgt atgagcatgc tgttgctcat
     3181 tctggattgc ctgacaatgc cagacgatcc ggtagcggtc acggtactgg ttctactggt
     3241 tctggttctt ctgagagtgc gattaagcca gacatgaaga tcaacctccg tatggaaggc
     3301 aacgtaaacg ggcaccactt tgtgatcgac ggagatggta caggcaagcc ttttgaggga
     3361 aaacagagta tggatcttga agtcaaagag ggcggacctc tgccttttgc ctttgatatc
     3421 ctgaccactg cattccatta cggcaacagg gtattcgccg aatatccaga ccacatacaa
     3481 gactatttta agcagtcgtt tcctaagggg tattcgtggg aacgaagctt gactttcgaa
     3541 gacgggggca tttgcattgc cagaaacgac ataacaatgg aaggggacac tttctataat
     3601 aaagttcgat ttcacggtgt aaactttccc gccaatggtc cagttatgca gaagaagacg
     3661 ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg atggagtgct gactggtgat
     3721 attcgcatgg ctttgttgct tgaaggaaat gcccattacc gatgtgactt cagaactact
     3781 tacaaagcta aggagaaggg tgtcaagtta ccaggctacc actttgtgga ccactgcatt
     3841 gagattttaa gccatgacaa agattacaac aaggttaagc tgtatgagca tgctgttgct
     3901 cattctggat tgcctgacaa tgccagacga taagcggccg cgactctaga tcataatcag
     3961 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4021 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     4081 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4141 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     4201 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4261 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4321 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4381 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4441 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4501 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4561 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4621 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4741 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4801 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4861 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4921 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4981 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5041 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5101 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     5161 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5221 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5281 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5341 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5401 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5461 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5521 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5581 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5641 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5701 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5761 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5821 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5881 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5941 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6001 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6061 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6121 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     6181 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6241 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6301 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6361 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6421 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6481 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7201 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7261 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     7321 agttccg
//
