LOCUS       sequence-192824-        5434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CETN1-iRFP670 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5434)
  AUTHORS   Bertolin G, Sizaire F, Herbomel G, Reboutier D, Prigent C, Tramier M
  TITLE     A FRET biosensor reveals spatiotemporal activation and functions of 
            aurora kinase A in living cells.
  JOURNAL   Nat Commun. 2016 Sep 14;7:12674. doi: 10.1038/ncomms12674.
  PUBMED    27624869
REFERENCE   2  (bases 1 to 5434)
  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: "2016-09-14"; volume: "7:12674. doi"; pages: " 
            10.1038/ncomms12674"
COMMENT     Sequence Label: pCETN1-IRFP670
FEATURES             Location/Qualifiers
     source          1..5434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     586..606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1232..2167
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    2289..2410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2326..2345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2380..2399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2417..2872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2554..2575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2766..2785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2899..3003
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3005..3362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3005..3196
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3028..3048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3213..3348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3397..4191
                     /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             3397..4191
                     /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(3451..3470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4061..4080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4379..4398)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4423..4470
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4799..5387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5288..5307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
       61 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      421 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      481 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      541 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      601 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      661 agcgctaccg gactcagatc tcgagctcaa gcttatggct tccggcttca agaagcccag
      721 cgctgcctcc accggccaaa agagaaaggt ggcacctaag cccgagctca ctgaggatca
      781 gaagcaagaa gttcgggaag catttgacct cttcgacgtg gacggaagtg ggaccatcga
      841 cgcgaaggag ctgaaggtgg ccatgagagc gctgggcttc gaacccagga aggaagagat
      901 gaagaaaatg atctccgagg tggacaggga aggcacgggg aagatcagct tcaatgactt
      961 cctggccgtg atgacgcaga agatgtccga gaaggacacc aaagaagaaa tcctgaaggc
     1021 cttcaggctc tttgatgacg atgagaccgg gaagatctcg ttcaaaaacc tgaagcgtgt
     1081 ggccaacgag ctgggggaga acctcacgga tgaggagctg caggagatga tcgacgaagc
     1141 tgatcgggat ggggacggcg aagtgaacga ggaggagttc cttcggatca tgaagaagac
     1201 cagcctttac ggggatccac cggtcgccac catggcgcgt aaggtcgatc tcacctcctg
     1261 cgatcgcgag ccgatccaca tccccggcag cattcagccg tgcggctgcc tgctagcctg
     1321 cgacgcgcag gcggtgcgga tcacgcgcat tacggaaaat gccggcgcgt tctttggacg
     1381 cgaaactccg cgggtcggtg agctactcgc cgattacttc ggcgagaccg aagcccatgc
     1441 gctgcgcaac gcactggcgc agtcctccga tccaaagcga ccggcgctga tcttcggttg
     1501 gcgcgacggc ctgaccggcc gcaccttcga catctcactg catcgccatg acggtacatc
     1561 gatcatcgag ttcgagcctg cggcggccga acaggccgac aatccgctgc ggctgacgcg
     1621 gcagatcatc gcgcgcacca aagaactgaa gtcgctcgaa gagatggccg cacgggtgcc
     1681 gcgctatctg caggcgatgc tcggctatca ccgcgtgatg ttgtaccgct tcgcggacga
     1741 cggctccggg atggtgatcg gcgaggcgaa gcgcagcgac ctcgagagct ttctcggtca
     1801 gcactttccg gcgtcgctgg tcccgcagca ggcgcggcta ctgtacttga agaacgcgat
     1861 ccgcgtggtc tcggattcgc gcggcatcag cagccggatc gtgcccgagc acgacgcctc
     1921 cggcgccgcg ctcgatctgt cgttcgcgca cctgcgcagc atctcgccct gccatctcga
     1981 atttctgcgg aacatgggcg tcagcgcctc gatgtcgctg tcgatcatca ttgacggcac
     2041 gctatgggga ttgatcatct gtcatcatta cgagccgcgt gccgtgccga tggcgcagcg
     2101 cgtcgcggcc gaaatgttcg ccgacttctt atcgctgcac ttcaccgccg cccaccacca
     2161 acgctaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2221 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2281 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2341 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2401 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2461 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2521 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2581 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2641 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2701 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2761 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2821 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     2881 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2941 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3001 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3181 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3241 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3301 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3361 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3421 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3481 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3541 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3601 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3661 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3721 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3781 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3841 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3901 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     3961 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4021 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4081 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4141 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4201 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4261 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4321 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4381 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4441 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4501 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4561 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4621 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4681 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4741 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5401 cggccttttt acggttcctg gccttttgct ggcc
//
