LOCUS       sequence-473506-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5955)
  TITLE     Chao Plasmids
REFERENCE   2  (bases 1 to 5955)
  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"
COMMENT     Sequence Label: p75-RFP
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    227..274
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     299..318
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(506..1300)
                     /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             complement(506..1300)
                     /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(617..636)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1227..1246
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1335..1692)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1349..1484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1501..1692
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1649..1669
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1694..1798)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1825..2280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1912..1931)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2122..2143
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2287..2408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2352..2371
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2525..3202)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             complement(2525..3202)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2587..2610)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2785..2804)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3054..3072
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(4583..4786)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4612..4632)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4787..5090
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5265..5853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5345..5364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag
       61 gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
      121 ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt
      181 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
      241 acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
      301 gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga
      361 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
      421 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
      481 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
      541 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
      601 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
      661 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
      721 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
      781 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
      841 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
      901 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
      961 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     1021 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     1081 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     1141 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     1201 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     1261 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     1321 tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     1381 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     1441 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     1501 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     1561 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     1621 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     1681 tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1801 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1861 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1921 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1981 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2041 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2101 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2161 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2221 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2281 acgccttaag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2341 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2401 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2461 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatctagagt
     2521 cgccttaggc gccggtggag tggcggccct cggcgcgctc gtactgttcc acgatggtgt
     2581 agtcctcgtt gtgggaggtg atgtccagct tgatgtcggt cttgtaggcg ccgggcagct
     2641 gcacgggctt cttggccatg taggtggtct tgacctcggc gtcgtagtgg ccgccgtcct
     2701 tcagcttcag cctcatcttg atctcgccct tcagggcgcc gtcctcgggg tacatccgct
     2761 cggtggaggc ctcccagccc atggtcttct tctgcattac ggggccgtcg gaggggaagt
     2821 tggtgccgcg cagcttcacc ttgtagatga actcgccgtc ctgcagggag gagtcctggg
     2881 tcacggtcac cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg
     2941 ggaaggacag cttcaagtag tcggggatgt cggcggggtg cttcacgtag gccttggagc
     3001 cgtactggaa ctgaggggac aggatgtccc aggcgaaggg cagggggccg cccttggtca
     3061 ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct
     3121 cgaactcgtg gccgttcacg gagccctcca tgcgcacctt gaagcgcatg aactccttga
     3181 tgacgtcctc ggaggaggcc atcggatccc gggcccggga tgtggcagtg gactcactgc
     3241 acagactctc cacgaggtcg gctcgctgga tgcggcgcag ggcggccagg agggcgtcca
     3301 gtgtggcgct gtcctgggtg gcccagcttg caagcagggc gcgaacgggg caggcctcat
     3361 gggtaaagga gtctatgtgc tcgggctggt agcccagctc gcccgccagg tgccgccagg
     3421 tgtcccccgc agagccgttg agaagcttct ccacctcctc ccgcttggct gggggcaggc
     3481 tgctgtagag gcctccgtca cccttgaggg cctggcccga ggctgtctgc gtgtggggct
     3541 gctggtcatg caggctctgg ctgtccacgg agatgccact gtcgctgtgg agtttttctc
     3601 cctctggtgg gggcgtctgg ttcactggcc ggctgttggc tccttgcttg ttctgcttgc
     3661 agctgttcca cctcttgaag gctatgtagg ccacaaggcc cacaaccaca gcagccagga
     3721 tggagcaata gacagggatg aggttgtcgg tggtgcctcg ggtcaccacg ggctgggagc
     3781 tgcccatcac tgtggtcacc acacctgcca ccgtgctggc tatgaggtct tgttctggag
     3841 gtgcctcagg ctcctgggtg ctgggggctg tgctgtccga gccctctggg ggtgtggacc
     3901 gtgtaatcca acggccaggg atctcctcgc actcggcgtc ggcccagcgt gtgcactcgc
     3961 ggagctggcg ctcggtgtcc tcgcacacgg tgcagggcag gcacgggtcc acgtggttgg
     4021 cctcgtcgga atacgtgccg tcggggcact cctcgcacac ggtgttctgc ttgtcctggc
     4081 aggagaacac gaggcccgag cccgcctcgc acacgcggca cgcctcgcag cgcccagtcg
     4141 tctcatcctg gtagtagccg taggcgcagc ggcacacggc gtcgtcggcc tccacgcacg
     4201 gcgccgacat gctctggagc cccacgcact cggtgcacgg cttgcacggc tcggtcgcgc
     4261 tcaccacgtc ggagaacgtc acgctgtcca ggcagggctc acacacggtc tggttggctc
     4321 cacaaggctg ggccacaccc tcgcccaggt tgcaggcttt gcagcactca ccgctgtgtg
     4381 tgtacaggcc tgtggggcat gcctccttgg cacctccaag ggacaccccc agaagcagca
     4441 acagcagcag gcgcggcccg tccatggcgc ggccggtggc acctgccccc atcgcccgcc
     4501 tcccgcggca gcgctcgaat tcgaagcttg agctcgagat ctgagtccgg tagcgctagc
     4561 ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     4621 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     4681 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     4741 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     4801 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4861 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4921 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4981 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     5041 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     5101 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg
     5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5941 aggatcttca cctag
//
