LOCUS       sequence-364934-        6845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labelled with near-infrared fluorescent protein 
            miRFP670nano3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6845)
  AUTHORS   Oliinyk OS, Baloban M, Clark CL, Carey E, Pletnev S, Nimmerjahn A, 
            Verkhusha VV
  TITLE     Single-domain near-infrared protein provides a scaffold for 
            antigen-dependent fluorescent nanobodies.
  JOURNAL   Nat Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6.
  PUBMED    35606446
REFERENCE   2  (bases 1 to 6845)
  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 
            Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6."
COMMENT     Sequence Label: pTubulin-miRFP670nano3
FEATURES             Location/Qualifiers
     source          1..6845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2768..2797
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2813..2830
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2853..2870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2859..3083
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3129..3557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3216..3235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3426..3447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3566..3586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3571..3900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3751..3886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3813..3832
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3948..3995
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4014..4388
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4518..4639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4555..4574)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4609..4628
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4688..4704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4688..4704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4701..4723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4712..4728
                     /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(4736..4742,4743..4760,4761..4766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4781..4802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4919..4936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5090..5678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5170..5189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5849..6640,6641..6709))
                     /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     6472..6491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6710..6814)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accatggcaa acctggacaa gatgctgaac accaccgtga ccgaggtgcg
      961 caagttcctg caagcggaca gagtgtgcgt gttcaagttc gaggaagatt actccggcac
     1021 cgtctcgcac gaagccgtgg acgacagatg gattagcatc ctgaagaccc aggtgcagga
     1081 cagatacttc atggaaacca gaggcgagga atacgtccac ggcagatacc aggccatcgc
     1141 cgacatctac acagccaatc tggtcgagtg ctacagagac ctgctgatcg agtttcaggt
     1201 gcgggccatt ctggctgtcc ccatcctgca aggcaagaag ctgtggggcc tgctggtggc
     1261 ccaccaactg gccggccctc gggagtggca gacctgggaa atcgacttcc tgaaacagca
     1321 agccgtggtg atgggcatcg ccatccagca gagcggaggt ggtggatcag gaggaagatc
     1381 tcgagtgcgt gagtgcatct ccatccacgt tggccaggct ggtgtccaga ttggcaatgc
     1441 ctgctgggag ctctactgcc tggaacacgg catccagccc gatggccaga tgccaagtga
     1501 caagaccatt gggggaggag atgactcctt caacaccttc ttcagtgaga cgggcgctgg
     1561 caagcacgtg ccccgggctg tgtttgtaga cttggaaccc acagtcattg atgaagttcg
     1621 cactggcacc taccgccagc tcttccaccc tgagcagctc atcacaggca aggaagatgc
     1681 tgccaataac tatgcccgag ggcactacac cattggcaag gagatcattg accttgtgtt
     1741 ggaccgaatt cgcaagctgg ctgaccagtg caccggtctt cagggcttct tggttttcca
     1801 cagctttggt gggggaactg gttctgggtt cacctccctg ctcatggaac gtctctcagt
     1861 tgattatggc aagaagtcca agctggagtt ctccatttac ccagcacccc aggtttccac
     1921 agctgtagtt gagccctaca actccatcct caccacccac accaccctgg agcactctga
     1981 ttgtgccttc atggtagaca atgaggccat ctatgacatc tgtcgtagaa acctcgatat
     2041 cgagcgccca acctacacta accttaaccg ccttattagc cagattgtgt cctccatcac
     2101 tgcttccctg agatttgatg gagccctgaa tgttgacctg acagaattcc agaccaacct
     2161 ggtgccctac ccccgcatcc acttccctct ggccacatat gcccctgtca tctctgctga
     2221 gaaagcctac catgaacagc tttctgtagc agagatcacc aatgcttgct ttgagccagc
     2281 caaccagatg gtgaaatgtg accctcgcca tggtaaatac atggcttgct gcctgttgta
     2341 ccgtggtgac gtggttccca aagatgtcaa tgctgccatt gccaccatca aaaccaagcg
     2401 cagcatccag tttgtggatt ggtgccccac tggcttcaag gttggcatca actaccagcc
     2461 tcccactgtg gtgcctggtg gagacctggc caaggtacag agagctgtgt gcatgctgag
     2521 caacaccaca gccattgctg aggcctgggc tcgcctggac cacaagtttg acctgatgta
     2581 tgccaagcgt gcctttgttc actggtacgt gggtgagggg atggaggaag gcgagttttc
     2641 agaggcccgt gaagatatgg ctgcccttga gaaggattat gaggaggttg gtgtggattc
     2701 tgttgaagga gagggtgagg aagaaggaga ggaatactaa ggatccaccg gatctagagg
     2761 gcccttcgaa caaaaactca tctcagaaga ggatctgaat atgcataccg gtcatcatca
     2821 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2881 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2941 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3001 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3061 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3121 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3181 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3241 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3301 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3361 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3421 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3481 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3541 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     3961 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     4021 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     4081 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     4141 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     4201 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     4261 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     4321 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     4381 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4441 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4501 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4561 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4621 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4681 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4741 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4801 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4861 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4921 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4981 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5041 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5101 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5161 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5221 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5281 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5341 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5401 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5461 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5521 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5581 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5641 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5701 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5761 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5821 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5881 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5941 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6001 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6061 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6121 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6181 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6241 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6301 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6361 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6421 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6481 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6541 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6601 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6661 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6721 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6781 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6841 acgtc
//
