LOCUS       sequence-140322-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse RFP-IKK-epsilon in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6756)
  TITLE     CAG.Luc.puro
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6756)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pRFP.mIKK-epsilon
FEATURES             Location/Qualifiers
     source          1..6756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(698..716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     966..985
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1160..1183
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3484..3605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3521..3540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3575..3594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3612..4067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3749..3770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3961..3980
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4094..4198
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4200..4557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4200..4391
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4223..4243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4408..4543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4592..5386
                     /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             4592..5386
                     /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(4646..4665)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5256..5275
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5574..5593)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5618..5665
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5994..6582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6483..6502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      601 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      661 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      721 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      781 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      901 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      961 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1021 cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc
     1081 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
     1141 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
     1201 gaacagtacg agcgcgccga gggccgccac tccaccggcg cctccggact cagatctatg
     1261 cagagtacca ctaactacct gtggcatact gatgacctgc tagggcaggg ggccactgcc
     1321 agtgtgtaca aggcccgaaa caagaaatcc ggggaggtgg ttgctgtaaa ggtcttcaac
     1381 tcagccagct atcggcgacc tcctgaggtt caggtgaggg agtttgaggt cctgcggagg
     1441 ctgaatcacc agaacatcgt gaagctattc gcagtggagg aaacgggagg cagccggcag
     1501 aaggtgctaa tcatggagta ctgctccagt gggagcctgc tgagcgtgct ggaagaccct
     1561 gagaacacgt tcgggctttc tgaagaggag ttcctagtgg tgctgcgctg tgtggtggct
     1621 ggcatgaacc acctgcggga gaatggcatt gtccatcggg acatcaaacc tgggaacatc
     1681 atgcgcctgg tgggcgagga ggggcagagc atctataagc tgtctgactt cggggctgcc
     1741 cgcaagctgg acgatgatga gaagtttgtt tctgtctatg gtacagagga atacctgcac
     1801 cctgacatgt atgagcgtgc agtgctgcgc aaaccccagc aaaaggcatt tggtgtgact
     1861 gtggatctct ggagtattgg ggtgaccctg taccacgcag ccacaggcag tctgcccttc
     1921 atccccttcg gtgggccccg gcgcaacaaa gagatcatgt acagaatcac cacagagaag
     1981 ccagccgggg ccatttcagg gactcagaag caggaaaatg gtcccttgga gtggagctac
     2041 agcctcccca tcacctgtag actgtccatg ggactgcaga accagctggt gcccatcctg
     2101 gccaacatcc tggaggtgga agaggataag tgctggggct ttgatcagtt cttcgcggag
     2161 accagtgaca ttctgcagcg aacggtcatc cacgtctttt ccctacccca ggccgttttg
     2221 catcatgtct acatccacgc ccacaacacg attgccatct ttttggaggc tgtatatgag
     2281 cagaccaacg tgacccccaa acaccaggag tacctcttcg agggtcaccc ttgtgtcctt
     2341 gagccaagcc tctcagccca gcacatcgcc cacacagctg ccagcagccc tctaactctg
     2401 ttcagcatgt ccagcgacac acctaagggg ctggccttca gggaccctgc tctggatgtc
     2461 ccaaagttcg tccctaaggt tgacctacag gccgattaca gcacagctaa gggggtgctg
     2521 ggcgctggct accaggccct gtggctggcg cgggtcctgc tggatggaca ggcgttgatg
     2581 cttcgggggt tacattgggt cctggaggtg cttcaggaca cgtgccagca gacactggag
     2641 gtcacacgga cagccctcct ctacctcagc agcagcctgg gcactgaaag gttcagcagt
     2701 ggagcgggga tgcctgacgt ccaggaacga aaggaggcca cagagctaag aaccaggctg
     2761 cagactctct cagagatcct gtctaaatgt tcccacaatg tcacagaaac ccaaaggagc
     2821 ctgagctgtc tgggtgaaga gcttttaaag aaccgggacc agattcatga ggataacaaa
     2881 agtatccaga agattcagtg ttgtttggac aagatgcact tcatctacaa acagttcaag
     2941 aaatccagga tgaggccagg gctcagctac aatgaggagc agatccacaa gctggataag
     3001 gtaaatttca gtcatctagc caagaggctg ctgcaggtgt tccaggagga gtgtgtgcag
     3061 acgtatcagg tgtcgctggt cacacacggc aagcggatga ggcaggtgca gagggcccag
     3121 aaccacctgc atctcattgg ccactctgtg gccacctgta actcggaagc ccggggagcc
     3181 caggagagtc tgaacaagat ctttgatcag ctccttctgg acagagcttc cgaacaggga
     3241 gctgaggtgt caccgcaacc tatggctcct catcccggcc ctgatccgaa ggacctggtc
     3301 ttccacatgc aggagctttg taagaattct gcagtcgacg gtaccgcggg cccgggatcc
     3361 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3421 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3481 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3601 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3661 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3721 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3781 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3841 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3901 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3961 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4021 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4081 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4141 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4201 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4321 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4381 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4441 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4501 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4561 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4621 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4681 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4741 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4801 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4861 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4921 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4981 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5041 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5101 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5161 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5221 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5281 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5341 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5401 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5461 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5521 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5581 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5641 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5701 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5761 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5821 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5881 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5941 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6121 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6661 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     6721 ggggtcatta gttcatagcc catatatgga gttccg
//
