LOCUS       sequence-410745-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labelled with near-infrared fluorescent protein 
            miRFP704nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  AUTHORS   Oliinyk OS, Pletnev S, Baloban M, Verkhusha VV
  TITLE     Development of bright red-shifted miRFP704nano using structural 
            analysis of miRFPnano proteins.
  JOURNAL   Protein Sci. 2023 Jun 22:e4709. doi: 10.1002/pro.4709.
  PUBMED    37347539
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: "Protein 
            Sci. 2023 Jun 22:e4709. doi: 10.1002/pro.4709."
COMMENT     Sequence Label: pTubulin-miRFP704nano
FEATURES             Location/Qualifiers
     source          1..6875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(120..140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        125..454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      305..440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     367..386
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        502..549
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             568..942
                     /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    1072..1193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1109..1128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1163..1182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1242..1258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1242..1258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1255..1277)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1266..1282
                     /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(1290..1296,1297..1314,1315..1320))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1335..1356
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1473..1490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1644..2232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1724..1743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2403..3194,3195..3263))
                     /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     3026..3045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3264..3368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3443..3462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3634..4013
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4014..4217
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4168..4188
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4262..4281
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4262..4280
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6197..6226
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6242..6259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6282..6299)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6288..6512
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6558..6875,1..111)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6645..6664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6855..6875,1)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
       61 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      121 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      181 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      361 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      421 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
      481 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
      541 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
      601 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
      661 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
      721 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
      781 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
      841 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
      901 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
      961 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     1021 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     1081 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1141 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1201 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1261 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1321 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1381 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1441 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1501 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1561 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1621 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1681 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1741 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1801 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1861 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1921 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1981 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2041 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2101 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2161 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2221 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2281 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2341 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2401 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2461 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2521 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2581 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2641 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2701 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2761 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2821 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2881 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2941 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3001 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3061 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3121 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3181 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3241 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3301 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3361 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     3421 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     3481 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     3541 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     3601 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     3661 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     3721 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     3781 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     3841 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     3901 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     3961 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     4021 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4081 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4141 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4201 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     4261 ttaatacgac tcactatagg gagacccaag ctggctagtt aagcttggta ccatggcaaa
     4321 cctggacaag atgctgaaca ccatcgtgac cgaggtgcgc cagttcctgc aagtggacag
     4381 agtgtgcgtg ttccagttcg aggaagatta ctccggcagc gtcgtcgtgg aagccgtgga
     4441 cgacagatgg aacagcatcc tgaagaccca ggtgcgggac tgctacttca tggaaaccag
     4501 aggtgaggaa tacttgcacg gcagatacca ggccatcgcc gacatctacc aggccaatct
     4561 gctggagagc tacagagacc tgctgggcca gtttcaggtg cgggccattc tggctgtccc
     4621 catcatcaag ggcaagaagc tgtggggcct gctggtggcc caccaactgg ctgcccctcg
     4681 gagctggcag acctgggaaa tcgagttcct gaaacagcaa gccgtggtga tgggcatcgc
     4741 catccagcag agcggaggtg gtggatcagg aggaggtggt tctggaggtg gtggatcagg
     4801 aggaagatct cgagtgcgtg agtgcatctc catccacgtt ggccaggctg gtgtccagat
     4861 tggcaatgcc tgctgggagc tctactgcct ggaacacggc atccagcccg atggccagat
     4921 gccaagtgac aagaccattg ggggaggaga tgactccttc aacaccttct tcagtgagac
     4981 gggcgctggc aagcacgtgc cccgggctgt gtttgtagac ttggaaccca cagtcattga
     5041 tgaagttcgc actggcacct accgccagct cttccaccct gagcagctca tcacaggcaa
     5101 ggaagatgct gccaataact atgcccgagg gcactacacc attggcaagg agatcattga
     5161 ccttgtgttg gaccgaattc gcaagctggc tgaccagtgc accggtcttc agggcttctt
     5221 ggttttccac agctttggtg ggggaactgg ttctgggttc acctccctgc tcatggaacg
     5281 tctctcagtt gattatggca agaagtccaa gctggagttc tccatttacc cagcacccca
     5341 ggtttccaca gctgtagttg agccctacaa ctccatcctc accacccaca ccaccctgga
     5401 gcactctgat tgtgccttca tggtagacaa tgaggccatc tatgacatct gtcgtagaaa
     5461 cctcgatatc gagcgcccaa cctacactaa ccttaaccgc cttattagcc agattgtgtc
     5521 ctccatcact gcttccctga gatttgatgg agccctgaat gttgacctga cagaattcca
     5581 gaccaacctg gtgccctacc cccgcatcca cttccctctg gccacatatg cccctgtcat
     5641 ctctgctgag aaagcctacc atgaacagct ttctgtagca gagatcacca atgcttgctt
     5701 tgagccagcc aaccagatgg tgaaatgtga ccctcgccat ggtaaataca tggcttgctg
     5761 cctgttgtac cgtggtgacg tggttcccaa agatgtcaat gctgccattg ccaccatcaa
     5821 aaccaagcgc agcatccagt ttgtggattg gtgccccact ggcttcaagg ttggcatcaa
     5881 ctaccagcct cccactgtgg tgcctggtgg agacctggcc aaggtacaga gagctgtgtg
     5941 catgctgagc aacaccacag ccattgctga ggcctgggct cgcctggacc acaagtttga
     6001 cctgatgtat gccaagcgtg cctttgttca ctggtacgtg ggtgagggga tggaggaagg
     6061 cgagttttca gaggcccgtg aagatatggc tgcccttgag aaggattatg aggaggttgg
     6121 tgtggattct gttgaaggag agggtgagga agaaggagag gaatactaag gatccaccgg
     6181 atctagaggg cccttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg
     6241 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     6301 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     6361 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     6421 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     6481 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     6541 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6601 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6781 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6841 cacgttcttt aatagtggac tcttgttcca aactg
//
