LOCUS       sequence-204548-        6297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Alpha-tubulin fused to monomeric near-infrared fluorescent protein 
            miRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 6297)
  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: "Nat Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pmiRFP-alpha-Tubulin-19-C
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3085..3206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3122..3141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3176..3195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3213..3668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3350..3371)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3562..3581
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3695..3799
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3801..4158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3801..3992
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3824..3844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4009..4144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4193..4987
                     /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             4193..4987
                     /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(4247..4266)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4857..4876
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5175..5194)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5219..5266
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5595..6183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6084..6103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccgcc
      601 accatggtcg caggacacgc aagtggaagc cccgactttg gaaccgccga tccgagcgac
      661 tgtgagagag aagaaatcca tctggccgga tccatccagc cacacggaac actgctggtg
      721 gtctccgagc ctgaccatag aatcattcag gcatctgcca acgccgctga attcctgaat
      781 ctgggaagtg tgctgggagt ccccctggct gagatcgacg gggatctgct gatcaagatt
      841 ctgccccacc tggaccctac cgcagaagga atgccagtgg ccgtcaggtg ccgcattgga
      901 aacccctcca cagagtacga tggactgatg catcgaccac ctgaaggagg actgatcatt
      961 gagctggaac gggctgggcc acccattgat ctgagcggaa cactggctcc cgcactggag
     1021 cgaattcgaa ctgcaggatc cctgagggct ctgtgcgacg atactgcact gctgttccag
     1081 cagtgtaccg gctacgaccg agtgatggtc tatcggttcg atgaacaggg gcacggagag
     1141 gtgtattcag aaatacatgt gactggactg gagagctact ttggcaacag gtatcctagc
     1201 tccctcgtgc cacagatggc ccggagactg tacgaaagac agagggtgcg cgtcctggtg
     1261 gatgtctctt atcagcctgt gccactggag ccaagactga gtcccctgac tggcagggac
     1321 ctggatatgt cagggtgttt tctgcgctct atgagtccta ctcacctgca gttcctgaag
     1381 aatatgggcg tgcgggcaac actggtggtc agcctggtgg tcggaggcaa actgtggggg
     1441 ctggtgatct gccaccatta tctgccacgc ttcatccatt ttgagctgcg agcaatttgt
     1501 gagctgctgg ccgaagctat cgcaacccgc attacagccc tgtccggagg cctcagatct
     1561 ggcagcggtg gaggcagcgc atccggcgga agcggaagcg tgcgtgagtg catctccatc
     1621 cacgttggcc aggctggtgt ccagattggc aatgcctgct gggagctcta ctgcctggaa
     1681 cacggcatcc agcccgatgg ccagatgcca agtgacaaga ccattggggg aggagatgac
     1741 tccttcaaca ccttcttcag tgagacgggc gctggcaagc acgtgccccg ggctgtgttt
     1801 gtagacttgg aacccacagt cattgatgaa gttcgcactg gcacctaccg ccagctcttc
     1861 caccctgagc agctcatcac aggcaaggaa gatgctgcca ataactatgc ccgagggcac
     1921 tacaccattg gcaaggagat cattgacctt gtgttggacc gaattcgcaa gctggctgac
     1981 cagtgcaccg gtcttcaggg cttcttggtt ttccacagct ttggtggggg aactggttct
     2041 gggttcacct ccctgctcat ggaacgtctc tcagttgatt atggcaagaa gtccaagctg
     2101 gagttctcca tttacccagc accccaggtt tccacagctg tagttgagcc ctacaactcc
     2161 atcctcacca cccacaccac cctggagcac tctgattgtg ccttcatggt agacaatgag
     2221 gccatctatg acatctgtcg tagaaacctc gatatcgagc gcccaaccta cactaacctt
     2281 aaccgcctta ttagccagat tgtgtcctcc atcactgctt ccctgagatt tgatggagcc
     2341 ctgaatgttg acctgacaga attccagacc aacctggtgc cctacccccg catccacttc
     2401 cctctggcca catatgcccc tgtcatctct gctgagaaag cctaccatga acagctttct
     2461 gtagcagaga tcaccaatgc ttgctttgag ccagccaacc agatggtgaa atgtgaccct
     2521 cgccatggta aatacatggc ttgctgcctg ttgtaccgtg gtgacgtggt tcccaaagat
     2581 gtcaatgctg ccattgccac catcaaaacc aagcgcagca tccagtttgt ggattggtgc
     2641 cccactggct tcaaggttgg catcaactac cagcctccca ctgtggtgcc tggtggagac
     2701 ctggccaagg tacagagagc tgtgtgcatg ctgagcaaca ccacagccat tgctgaggcc
     2761 tgggctcgcc tggaccacaa gtttgacctg atgtatgcca agcgtgcctt tgttcactgg
     2821 tacgtgggtg aggggatgga ggaaggcgag ttttcagagg cccgtgaaga tatggctgcc
     2881 cttgagaagg attatgagga ggttggtgtg gattctgttg aaggagaggg tgaggaagaa
     2941 ggagaggaat actaaggatc caccggatct agataactga tcataatcag ccataccaca
     3001 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3061 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3121 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3181 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     3241 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3301 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3361 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3421 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3481 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3541 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3601 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3661 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3721 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3781 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3841 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3901 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4021 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4081 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4141 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4201 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4261 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4321 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4381 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4441 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4501 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4561 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4621 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4681 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4741 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4801 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4861 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4921 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4981 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5041 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5101 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5161 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5221 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5281 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5341 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5401 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5461 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5701 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6241 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
