LOCUS       sequence-163479-        6123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PAsfCherry2(1-10) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6123)
  AUTHORS   Feng S, Sekine S, Pessino V, Li H, Leonetti MD, Huang B
  TITLE     Improved split fluorescent proteins for endogenous protein labeling.
  JOURNAL   Nat Commun. 2017 Aug 29;8(1):370. doi: 10.1038/s41467-017-00494-8.
  PUBMED    28851864
REFERENCE   2  (bases 1 to 6123)
  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: "2017-08-29"; volume: "8(1):370. doi"; pages: " 
            10.1038/s41467-017-00494-8"
COMMENT     Sequence Label: pcDNA3.1_PAsfCherry2(1-10)
FEATURES             Location/Qualifiers
     source          1..6123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1056..1074)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1324..1343
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1627..1656
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1672..1689
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1712..1729)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1718..1942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1988..2416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2075..2094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2285..2306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2425..2445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2430..2759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2610..2745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2672..2691
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2826..3620
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2880..2899)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3490..3509
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3796..3917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3833..3852)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3887..3906
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3966..3982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3966..3982)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3979..4001)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3990..4006
                     /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(4014..4020,4021..4038,4039..4044))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4059..4080
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4197..4214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4368..4956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4448..4467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5127..5918,5919..5987))
                     /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     5750..5769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5988..6092)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccatggagga ggataacatg gccatcatta aggagttcat
      961 gcgcttcaag gtgcacatgg aggggtccgt gaacggccac gtgttcgaga tcgagggcga
     1021 gggcgagggc cacccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     1081 ccccctgccc ttcacctggg acatcctgtc ccctcaattc atgtacggct ccaatgccta
     1141 cgtgaagcac cccgccgaca tccccgacta ctttaagctg tccttccccg agggcttcac
     1201 ctgggagcgc gtgatgaaat tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1261 cctgcaggac ggtcagttca tctacaaggt gaagctgctg ggcatcaact tcccctccga
     1321 cggccccgta atgcagaaga agaccatggg ctgggaggcc ctcaccgagc ggatgtaccc
     1381 cgaggacggc gccctgaagg gcgaggtcaa cccgagagtg aagctgaagg acggcggcca
     1441 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     1501 ctacaacgtc gaccgcaagt tggacatcac ctcacacaac gaggactgag gatccactag
     1561 tccagtgtgg tggaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg
     1621 cccttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg tcatcatcac
     1681 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1741 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1801 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1861 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     1921 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     1981 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2041 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2101 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2161 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2221 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2281 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2341 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2401 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2761 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     2821 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2881 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2941 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3001 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3061 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3121 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3181 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3241 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3301 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3361 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3421 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3481 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3541 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3601 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     3661 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3721 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3781 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3841 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3901 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3961 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4021 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4081 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4141 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4201 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4261 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4321 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4381 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4441 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4501 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4561 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4621 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4681 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4741 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4801 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4861 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4921 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4981 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5041 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5101 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5161 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5221 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5281 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5341 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5401 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5461 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5521 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5581 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5641 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5701 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5761 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5821 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5881 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5941 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6061 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6121 gtc
//
