LOCUS       sequence-271530-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zyxin labeling with near-infrared fluorescent protein emiRFP703 
            (enhanced miRFP703).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  AUTHORS   Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti 
            F, Shemetov AA, Testa I, Verkhusha VV
  TITLE     A set of monomeric near-infrared fluorescent proteins for multicolor
            imaging across scales.
  JOURNAL   Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
  PUBMED    31932632
REFERENCE   2  (bases 1 to 6636)
  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: "2020-01-13"; volume: "11(1):239. doi"; pages: " 
            10.1038/s41467-019-13897-6"
COMMENT     Sequence Label: pzyxin-emiRFP703
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2382..2391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3442..3563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3479..3498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3533..3552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3570..4025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3707..3728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3919..3938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4052..4156
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4158..4515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4158..4349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4181..4201)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4366..4501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4550..5344
                     /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             4550..5344
                     /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(4604..4623)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5214..5233
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5532..5551)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5576..5623
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5952..6540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6441..6460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      601 tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat tccatggcgg
      661 ccccccgccc gtctcccgcg atccccgttt cggtctcggc tccggctttt tacgccccgc
      721 agaagaagtt cggccctgtg gtggccccaa agcccaaagt gaatcccttc cggcccaggg
      781 acagcgagcc tcccccggca cccggggccc agcgcgcaca gatgggccgg gtgggcgata
      841 ttcccccgcc gcccccggaa gactttcccc tgcctccacc tccccttgct ggggatggcg
      901 acgatgcaga gggtgctctg ggaggtgcct tcccgccgcc ccctcccccg atcgaggaat
      961 catttccccc tgcgcctctg gaggaggaga tcttcccttc cccgccgcct cctccggagg
     1021 aggagggagg gcctgaggcc cccataccgc ccccaccaca gcccagggag aaggtgagca
     1081 gtattgattt ggagatcgac tctctgtcct cactgctgga tgacatgacc aagaatgatc
     1141 ctttcaaagc ccgggtgtca tctggatatg tgcccccacc agtggccact ccattcagtt
     1201 ccaagtccag taccaagcct gcagccgggg gcacagcacc cctgcctcct tggaagtccc
     1261 cttccagctc ccagcctctg ccccaggttc cggctccggc tcagagccag acacagttcc
     1321 atgttcagcc ccagccccag cccaagcctc aggtccaact ccatgtccag tcccagaccc
     1381 agcctgtgtc tttggctaac acccagcccc gagggccccc agcctcatct ccggctccag
     1441 cccctaagtt ttctccagtg actcctaagt ttactcctgt ggcttccaag ttcagtcctg
     1501 gagccccagg tggatctggg tcacaaccaa atcaaaaatt ggggcacccc gaagctcttt
     1561 ctgctggcac aggctcccct caacctccca gcttcaccta tgcccagcag agggagaagc
     1621 cccgagtgca ggagaagcag caccccgtgc ccccaccggc tcagaaccaa aaccaggtgc
     1681 gctcccctgg ggccccaggg cccctgactc tgaaggaggt ggaggagctg gagcagctga
     1741 cccagcagct aatgcaggac atggagcatc ctcagaggca gaatgtggct gtcaacgaac
     1801 tctgcggccg atgccatcaa cccctggccc gggcgcagcc agccgtccgc gctctagggc
     1861 agctgttcca catcgcctgc ttcacctgcc accagtgtgc gcagcagctc cagggccagc
     1921 agttctacag tctggagggg gcgccgtact gcgagggctg ttacactgac accctggaga
     1981 agtgtaacac ctgcggggag cccatcactg accgcatgct gagggccacg ggcaaggcct
     2041 atcacccgca ctgcttcacc tgtgtggtct gcgcccgccc cctggagggc acctccttca
     2101 tcgtggacca ggccaaccgg ccccactgtg tccccgacta ccacaagcag tacgccccga
     2161 ggtgctccgt ctgctctgag cccatcatgc ctgagcctgg ccgagatgag actgtgcgag
     2221 tggtcgccct ggacaagaac ttccacatga agtgttacaa gtgtgaggac tgcgggaagc
     2281 ccctgtcgat tgaggcagat gacaatggct gcttccccct ggacggtcac gtgctctgtc
     2341 ggaagtgcca cactgctaga gcccagacgg atccaccggt cgccaccatg gcggaaggct
     2401 ccgtcgccag gcagcctgac ctcttgacct gcgaacatga agagatccac ctcgccggct
     2461 cgatccagcc gcatggcgcg cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg
     2521 ccagcgccaa cgccgcggaa tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg
     2581 agatcgacgg cgatctgttg atcaagatcc tgccgcatct cgatcccacc gccgaaggca
     2641 tgccggtcgc ggtgcgctgc cggatcggca atccctctac ggagtactgc ggtctgatgc
     2701 atcggcctcc ggaaggcggg ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc
     2761 tgtcaggcac gctggcgccg gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc
     2821 tgtgcgatga caccgtgctg ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt
     2881 atcgtttcga tgagcaaggc cacggcctgg tattctccga gtgccatgtg cctgggctcg
     2941 aatcctattt cggcaaccgc tatccgtcgt cgctggtccc gcagatggcg cggcagctgt
     3001 acgtgcggca gcgcgtccgc gtgctggtcg acgtcaccta tcagccggtg ccgctggagc
     3061 cgcggctgtc gccgctgacc gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga
     3121 tgtcgccgat ccatctgcag ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt
     3181 cgctggtggt cggcggcaag ctgtggggcc tggttgtctg tcaccattat ctgccgcgct
     3241 tcatccgttt cgagctgcgg gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga
     3301 tcaccgcgct tgagagctaa gcggccgcga ctctagatca taatcagcca taccacattt
     3361 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3421 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3541 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     3601 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3661 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3721 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3781 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3841 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3901 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3961 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4021 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4141 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4321 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4381 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4441 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4501 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4561 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4621 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4681 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4741 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4801 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4861 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4921 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4981 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5041 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5101 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5161 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5221 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5281 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5341 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5401 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5461 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5521 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5581 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5641 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5701 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5761 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5821 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5881 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5941 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6001 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6061 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6121 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6181 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6241 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6301 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6361 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6421 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6481 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6541 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6601 ttctttcctg cgttatcccc tgattctgtg gataac
//
