LOCUS       sequence-384903-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labelled with near-infrared fluorescent protein 
            miRFP718nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  AUTHORS   Oliinyk OS, Ma C, Pletnev S, Baloban M, Toboada C, Sheng H, Yao J, 
            Verkhusha VV
  TITLE     Deep-tissue SWIR imaging using rationally designed small red-shifted
            near-infrared fluorescent protein.
  JOURNAL   Nat Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0.
  PUBMED    36456785
REFERENCE   2  (bases 1 to 6875)
  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 Dec 1. doi: 10.1038/s41592-022-01683-0."
COMMENT     Sequence Label: pTubulin-miRFP718nano
FEATURES             Location/Qualifiers
     source          1..6875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     780..799
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        780..798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2715..2744
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2760..2777
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2800..2817)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2806..3030
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3076..3504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3163..3182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3373..3394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3513..3533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3518..3847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3698..3833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3760..3779
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3895..3942
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3961..4335
                     /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    4465..4586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4502..4521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4556..4575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4635..4651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4635..4651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4648..4670)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4659..4675
                     /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(4683..4689,4690..4707,4708..4713))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4728..4749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4866..4883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5037..5625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5117..5136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5796..6587,6588..6656))
                     /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     6419..6438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6657..6761)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6836..6855)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
       61 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      121 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
      781 aatacgactc actataggga gacccaagct ggctagttaa gcttggtacc atggcaaacc
      841 tggacaagat gctgaacacc atcgtgaccg aggtgcgcca gttcctgcaa gtggacagac
      901 tctgcgtgtt caagttcgag gaagattact ccggcaacat catctacgaa gccgtggacg
      961 acggatggtt gagcatcctg aagacccacg tgcgggactg ctacttcatg gaaaccagag
     1021 gcgaggaata cctgcacggc agataccagg ccatcgccga catccaccag gccaatctgg
     1081 cggagagcta tagagacttc ctgacccagt accaggtgcg ggccattgtg gctgtcccca
     1141 tcctgaaggg caagaagctg tggggcttgt tcagcgccca ccaactggcc gcccctcgga
     1201 gctggcaggc ctgggaaatc gagttcctga aacagcaagc cgtggtgatg ggcatcgcca
     1261 tccagcagag cggaggtggt ggatcaggag gaggtggttc tggaggtggt ggatcaggag
     1321 gaagatctcg agtgcgtgag tgcatctcca tccacgttgg ccaggctggt gtccagattg
     1381 gcaatgcctg ctgggagctc tactgcctgg aacacggcat ccagcccgat ggccagatgc
     1441 caagtgacaa gaccattggg ggaggagatg actccttcaa caccttcttc agtgagacgg
     1501 gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt ggaacccaca gtcattgatg
     1561 aagttcgcac tggcacctac cgccagctct tccaccctga gcagctcatc acaggcaagg
     1621 aagatgctgc caataactat gcccgagggc actacaccat tggcaaggag atcattgacc
     1681 ttgtgttgga ccgaattcgc aagctggctg accagtgcac cggtcttcag ggcttcttgg
     1741 ttttccacag ctttggtggg ggaactggtt ctgggttcac ctccctgctc atggaacgtc
     1801 tctcagttga ttatggcaag aagtccaagc tggagttctc catttaccca gcaccccagg
     1861 tttccacagc tgtagttgag ccctacaact ccatcctcac cacccacacc accctggagc
     1921 actctgattg tgccttcatg gtagacaatg aggccatcta tgacatctgt cgtagaaacc
     1981 tcgatatcga gcgcccaacc tacactaacc ttaaccgcct tattagccag attgtgtcct
     2041 ccatcactgc ttccctgaga tttgatggag ccctgaatgt tgacctgaca gaattccaga
     2101 ccaacctggt gccctacccc cgcatccact tccctctggc cacatatgcc cctgtcatct
     2161 ctgctgagaa agcctaccat gaacagcttt ctgtagcaga gatcaccaat gcttgctttg
     2221 agccagccaa ccagatggtg aaatgtgacc ctcgccatgg taaatacatg gcttgctgcc
     2281 tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc tgccattgcc accatcaaaa
     2341 ccaagcgcag catccagttt gtggattggt gccccactgg cttcaaggtt ggcatcaact
     2401 accagcctcc cactgtggtg cctggtggag acctggccaa ggtacagaga gctgtgtgca
     2461 tgctgagcaa caccacagcc attgctgagg cctgggctcg cctggaccac aagtttgacc
     2521 tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg tgaggggatg gaggaaggcg
     2581 agttttcaga ggcccgtgaa gatatggctg cccttgagaa ggattatgag gaggttggtg
     2641 tggattctgt tgaaggagag ggtgaggaag aaggagagga atactaagga tccaccggat
     2701 ctagagggcc cttcgaacaa aaactcatct cagaagagga tctgaatatg cataccggtc
     2761 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2821 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2881 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2941 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3001 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3061 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3121 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3181 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3241 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3301 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3361 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3421 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3481 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3601 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3781 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3841 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     3901 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     3961 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     4021 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     4081 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     4141 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     4201 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     4261 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     4321 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4381 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4441 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4501 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4561 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4621 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4681 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4741 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4801 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4861 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4921 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4981 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5041 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5101 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5161 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5221 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5281 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5341 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5401 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5461 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5521 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5581 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5641 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5701 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5761 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5821 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5881 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5941 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6001 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6061 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6121 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6181 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6241 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6301 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6361 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6421 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6481 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6541 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6601 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6661 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6721 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6781 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6841 tctgctctga tgccgcatag ttaagccagt atctg
//
