LOCUS       sequence-388156-        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion protein of Cx43-7-miRFP670-2 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  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 6071)
  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-miRFP670-2-N1
FEATURES             Location/Qualifiers
     source          1..6071
                     /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..2737
                     /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
                     PRAVPMAQRVAAKRFAERLSTHFTAAHHQR"
     polyA_signal    2859..2980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2896..2915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2950..2969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2987..3442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3124..3145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3336..3355
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3469..3573
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3575..3932
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3575..3766
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3598..3618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3783..3918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3967..4761
                     /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             3967..4761
                     /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(4021..4040)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4631..4650
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4949..4968)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4993..5040
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5369..5957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5858..5877
                     /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 catggcgcgt aaggtcgatc tcacctcctg cgatcgcgag ccgatccaca tccccggcag
     1861 cattcagccg tgcggctgcc tgctagcctg cgacgcgcag gcggtgcgga tcacgcgcat
     1921 tacggaaaat gccggcgcgt tctttggacg cgaaactccg cgggtcggtg agctactcgc
     1981 cgattacttc ggcgagaccg aagcccatgc gctgcgcaac gcactggcgc agtcctccga
     2041 tccaaagcga ccggcgctga tcttcggttg gcgcgacggc ctgaccggcc gcaccttcga
     2101 catctcactg catcgccatg acggtacatc gatcatcgag ttcgagcctg cggcggccga
     2161 acaggccgac aatccgctgc ggctgacgcg gcagatcatc gcgcgcacca aagaactgaa
     2221 gtcgctcgaa gagatggccg cacgggtgcc gcgctatctg caggcgatgc tcggctatca
     2281 ccgcgtgatg ttgtaccgct tcgcggacga cggctccggg atggtgatcg gcgaggcgaa
     2341 gcgcagcgac ctcgagagct ttctcggtca gcactttccg gcgtcgctgg tcccgcagca
     2401 ggcgcggcta ctgtacttga agaacgcgat ccgcgtggtc tcggattcgc gcggcatcag
     2461 cagccggatc gtgcccgagc acgacgcctc cggcgccgcg ctcgatctgt cgttcgcgca
     2521 cctgcgcagc atctcgccct gccatctcga atttctgcgg aacatgggcg tcagcgcctc
     2581 gatgtcgctg tcgatcatca ttgacggcac gctatgggga ttgatcatct gtcatcatta
     2641 cgagccgcgt gccgtgccga tggcgcagcg cgtcgcggcc aaaaggttcg ccgagcgctt
     2701 atcgacgcac ttcaccgccg cccaccacca acgctaagcg gccgcgactc tagatcataa
     2761 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2821 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2881 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2941 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     3001 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3061 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3121 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3181 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3241 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3301 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3361 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3421 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3481 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3541 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3601 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3661 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3781 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3841 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3901 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3961 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4021 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4081 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4141 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4201 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4261 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4321 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4381 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4441 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4501 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4561 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4621 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4681 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4741 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4801 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4861 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4921 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4981 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5041 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5101 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5161 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5221 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5281 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5341 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5401 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5461 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5521 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgact acatacctcg
     5581 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5641 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5701 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5761 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5821 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5881 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5941 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6001 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6061 ccgccatgca t
//
