LOCUS       sequence-250232-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labeling with near-infrared fluorescent protein 
            miRFP670nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 7232)
  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 
            Commun."; date: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
COMMENT     Sequence Label: pTubulin-miRFP670nano
FEATURES             Location/Qualifiers
     source          1..7232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        137..516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     671..691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     765..784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        765..783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2687..2705)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2688..2705)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2725..2742)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2731..2955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3001..3429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3088..3107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3298..3319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3438..3458)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3443..3772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3623..3758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3685..3704
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3839..4633
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3893..3912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4503..4522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4807..4928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4844..4863)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4898..4917
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4977..4993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4977..4993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4990..5012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5001..5017
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5025..5031,5032..5049,5050..5055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5070..5091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5208..5225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5379..5967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5459..5478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6138..6929,6930..6998))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6761..6780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6999..7103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7178..7197)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
       61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      781 agggagaccc aagcttggta ccatggcaaa cctggacaag atgctgaata ccacagtaac
      841 agaggtgcgg cagttcctgc aggtggacag agtgtgcgtg ttccagtttg aggaggatta
      901 tagcggagtg gtggtggtgg aggccgtgga cgataggtgg atctccatcc tgaagaccca
      961 ggtgcgggat agatacttca tggagacaag gggcgaggag tattctcacg gccgctacca
     1021 ggccatcgcc gacatctaca ccgcaaacct gacagagtgc tacagggatc tgctgacaca
     1081 gtttcaggtg agagcaatcc tggccgtgcc catcctgcag ggcaagaagc tgtggggcct
     1141 gttggtggca caccagctgg cggcccctag acagtggcag acctgggaga tcgactttct
     1201 gaagcagcag gccgtggtgg tgggcatcgc catccagcag agcggaggtg gtggatcagg
     1261 aggaggtggt tctggaggtg gtggatcagg aggaagatct cgagtgcgtg agtgcatctc
     1321 catccacgtt ggccaggctg gtgtccagat tggcaatgcc tgctgggagc tctactgcct
     1381 ggaacacggc atccagcccg atggccagat gccaagtgac aagaccattg ggggaggaga
     1441 tgactccttc aacaccttct tcagtgagac gggcgctggc aagcacgtgc cccgggctgt
     1501 gtttgtagac ttggaaccca cagtcattga tgaagttcgc actggcacct accgccagct
     1561 cttccaccct gagcagctca tcacaggcaa ggaagatgct gccaataact atgcccgagg
     1621 gcactacacc attggcaagg agatcattga ccttgtgttg gaccgaattc gcaagctggc
     1681 tgaccagtgc accggtcttc agggcttctt ggttttccac agctttggtg ggggaactgg
     1741 ttctgggttc acctccctgc tcatggaacg tctctcagtt gattatggca agaagtccaa
     1801 gctggagttc tccatttacc cagcacccca ggtttccaca gctgtagttg agccctacaa
     1861 ctccatcctc accacccaca ccaccctgga gcactctgat tgtgccttca tggtagacaa
     1921 tgaggccatc tatgacatct gtcgtagaaa cctcgatatc gagcgcccaa cctacactaa
     1981 ccttaaccgc cttattagcc agattgtgtc ctccatcact gcttccctga gatttgatgg
     2041 agccctgaat gttgacctga cagaattcca gaccaacctg gtgccctacc cccgcatcca
     2101 cttccctctg gccacatatg cccctgtcat ctctgctgag aaagcctacc atgaacagct
     2161 ttctgtagca gagatcacca atgcttgctt tgagccagcc aaccagatgg tgaaatgtga
     2221 ccctcgccat ggtaaataca tggcttgctg cctgttgtac cgtggtgacg tggttcccaa
     2281 agatgtcaat gctgccattg ccaccatcaa aaccaagcgc agcatccagt ttgtggattg
     2341 gtgccccact ggcttcaagg ttggcatcaa ctaccagcct cccactgtgg tgcctggtgg
     2401 agacctggcc aaggtacaga gagctgtgtg catgctgagc aacaccacag ccattgctga
     2461 ggcctgggct cgcctggacc acaagtttga cctgatgtat gccaagcgtg cctttgttca
     2521 ctggtacgtg ggtgagggga tggaggaagg cgagttttca gaggcccgtg aagatatggc
     2581 tgcccttgag aaggattatg aggaggttgg tgtggattct gttgaaggag agggtgagga
     2641 agaaggagag gaatactaag gatccaccgg atctagaggg ccctattcta tagtgtcacc
     2701 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2761 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2821 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2881 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2941 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3001 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3061 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3421 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3481 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3541 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3601 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3661 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3721 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3781 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3841 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3901 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3961 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4021 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4081 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4141 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4201 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4261 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4321 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4381 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4441 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4501 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4561 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4621 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4681 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4741 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4801 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4861 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4921 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4981 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5041 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5101 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5221 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5281 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5521 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6061 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6121 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6181 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6241 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6301 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6361 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6421 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6481 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6541 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6601 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6661 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6721 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6781 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6841 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6901 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6961 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7021 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7081 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7141 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     7201 agttaagcca gtatctgctc cctgcttgtg tg
//
