LOCUS       sequence-388145-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion protein of Cx43-7-miRFP720 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 6086)
  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. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pCx43-7-miRFP720-N1
FEATURES             Location/Qualifiers
     source          1..6086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1796..1805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1802..2752
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    2874..2995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2911..2930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2965..2984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3002..3457)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3139..3160)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3351..3370
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3484..3588
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3590..3947
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3590..3781
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3613..3633)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3798..3933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3982..4776
                     /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             3982..4776
                     /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(4036..4055)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4646..4665
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4964..4983)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5008..5055
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5384..5972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5873..5892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctactac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatgggt gactggagtg ccttggggaa
      661 gcttctggac aaggtccaag cctactccac cgctggaggg aaggtgtggc tgtcagtgct
      721 cttcatattc agaatcctgc tcctggggac agctgttgag tcagcttggg gtgatgaaca
      781 gtctgccttt cgctgtaaca ctcaacaacc tggctgcgaa aacgtctgct atgacaagtc
      841 cttccccatc tctcacgtgc gcttctgggt ccttcagatc atattcgtgt ctgtgcccac
      901 actcctgtac ttggcccatg tgttctatgt gatgaggaag gaagagaagc taaacaagaa
      961 agaagaggag ctcaaagtgg cccagactga cggggtcaac gtggagatgc acctgaagca
     1021 gattgaaatc aagaagttca agtacgggat tgaagagcac ggcaaggtga aaatgagggg
     1081 cggcttgctg agaacctaca tcatcagcat cctcttcaag tctgtcttcg aggtggcctt
     1141 cctgctcatc cagtggtaca tctatgggtt cagcttgagc gcggtctaca cctgcaagag
     1201 agatccctgc ccccaccagg tagactgctt cctctcacgt cccacggaga aaaccatctt
     1261 catcatcttc atgctggtgg tgtccttggt gtctctcgct ttgaacatca ttgagctctt
     1321 ctacgtcttc ttcaagggcg ttaaggatcg cgtgaaggga agaagcgatc cttaccacgc
     1381 caccactggc ccactgagcc catcaaaaga ctgcggatct ccaaaatacg cctacttcaa
     1441 tggctgctcc tcaccaacgg ctccactctc gcctatgtct cctcctgggt acaagctggt
     1501 tactggtgac agaaacaatt cctcgtgccg caattacaac aagcaagcta gcgagcaaaa
     1561 ctgggcgaac tacagcgcag agcaaaatcg catggggcag gccggaagca ccatctccaa
     1621 ctcgcacgcc cagccgttcg atttccccga cgacaaccag aatgccaaaa aagttgctgc
     1681 tggacatgaa ctccagccat tagccatcgt ggaccaacga ccttccagca gagccagcag
     1741 ccgcgccagc agcaggcctc ggcctgatga cctggagatt gcggatccac cggtcgccac
     1801 catggcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg atgagccgat
     1861 ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg ccgacatgac
     1921 gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg gcgccctgat
     1981 cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga cgatcgcctt
     2041 ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga tgcgaaagga
     2101 cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg agctcgagcc
     2161 tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca gcgccatccg
     2221 ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc aagaggtgcg
     2281 gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact tcagcgggtc
     2341 cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc actatcctgc
     2401 ctcattcatc ccggcgcagg cccgtcggct ctataccatc aacccggtac ggatcattcc
     2461 cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca ccgggcggcc
     2521 gattgatctt agcttcgcca tcctgcgcag cgtctcgccc aaccatctgg agttcatgcg
     2581 caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc gactgtgggg
     2641 attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc gccaagcctg
     2701 caagagagtc gccgagagac tggccaccca gatcggcgtg atggaagagt aagcggccgc
     2761 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2821 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3001 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3061 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3121 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3181 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3241 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3301 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3361 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3421 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3601 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3781 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3841 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3901 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3961 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4021 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4081 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4141 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4201 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4261 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4321 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4381 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4441 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4501 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4561 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4621 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4681 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4741 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4801 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4861 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4921 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4981 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5041 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5101 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5161 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5221 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5401 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5461 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5521 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5581 cgactacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5641 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5701 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5761 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5821 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5881 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5941 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6001 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6061 tggataaccg tattaccgcc atgcat
//
