LOCUS       sequence-242400-        8181 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 8181)
  AUTHORS   Mendell JT, ap Rhys CM, Dietz HC
  TITLE     Separable roles for rent1/hUpf1 in altered splicing and decay of 
            nonsense transcripts.
  JOURNAL   Science. 2002 Oct 11. 298(5592):419-22.
  PUBMED    12228722
REFERENCE   2  (bases 1 to 8181)
  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: "Science. 
            2002 Oct 11. 298(5592):419-22."
COMMENT     Sequence Label: RNT1-GFP
FEATURES             Location/Qualifiers
     source          1..8181
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4969..5090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5006..5025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5060..5079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5097..5552)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5234..5255)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5446..5465
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5579..5683
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5685..6042
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5685..5876
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5708..5728)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5893..6028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6077..6871
                     /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             6077..6871
                     /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(6131..6150)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6741..6760
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7059..7078)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7103..7150
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7479..8067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7968..7987
                     /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 tcgctattac
      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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagtg cagcgcggaa ccggcccgag ggccctaccc
     1381 ggaggcacca tgagcgtgga ggcgtacggg cccagctcgc agactctcac tttcctggac
     1441 acggaggagg ccgagctgct tggcgccgac acacagggct ccgagttcga gttcaccgac
     1501 tttactcttc ctagccagac gcagacgccc cccggcggcc ccggcggccc gggcggtggc
     1561 ggcgcgggag gcccgggcgg cgcgggcgcg ggcgctgcgg cgggacagct cgacgcgcag
     1621 gttgggcccg aaggcatcct gcagaacggg gctgtggacg acagtgtagc caagaccagc
     1681 cagttgttgg ctgagttgaa cttcgaggaa gatgaagaag acacctatta cacgaaggac
     1741 ctccccatac acgcctgcag ttactgtgga atacacgatc ctgcctgcgt ggtttactgt
     1801 aataccagca agaagtggtt ctgcaacgga cgtggaaata cttctggcag ccacattgta
     1861 aatcaccttg tgagggcaaa atgcaaagag gtgaccctgc acaaggacgg gcccctgggg
     1921 gagacagtcc tggagtgcta caactgcggc tgtcgcaacg tcttcctcct cggcttcatc
     1981 ccggccaaag ctgactcagt ggtggtgctg ctgtgcaggc agccctgtgc cagccagagc
     2041 agcctcaagg acatcaactg ggacagctcg cagtggcagc cgctgatcca ggaccgctgc
     2101 ttcctgtcct ggctggtcaa gatcccctcc gagcaggagc agctgcgggc acgccagatc
     2161 acggcacagc agatcaacaa gctggaggag ctgtggaagg aaaacccttc tgccacgctg
     2221 gaggacctgg agaagccggg ggtggacgag gagccgcagc atgtcctcct gcggtacgag
     2281 gacgcctacc agtaccagaa catattcggg cccctggtca agctggaggc cgactacgac
     2341 aagaagctga aggagtccca gactcaagat aacatcactg tcaggtggga cctgggcctt
     2401 aacaagaaga gaatcgccta cttcactttg cccaagactg actctgacat gcggctcatg
     2461 cagggggatg agatatgcct gcggtacaaa ggggaccttg cgcccctgtg gaaagggatc
     2521 ggccacgtca tcaaggtccc tgataattat ggcgatgaga tcgccattga gctgcggagc
     2581 agcgtgggtg cacctgtgga ggtgactcac aacttccagg tggattttgt gtggaagtcg
     2641 acctcctttg acaggatgca gagcgcattg aaaacgtttg ccgtggatga gacctcggtg
     2701 tctggctaca tctaccacaa gctgttgggc cacgaggtgg aggacgtaat catcaagtgc
     2761 cagctgccca agcgcttcac ggcgcagggc ctccccgacc tcaaccactc ccaggtttat
     2821 gccgtgaaga ctgtgctgca aagaccactg agcctgatcc agggcccgcc aggcacgggg
     2881 aagacggtga cgtcggccac catcgtctac cacctggccc ggcaaggcaa cgggccggtg
     2941 ctggtgtgtg ctccgagcaa catcgccgtg gaccagctaa cggagaagat ccaccagacg
     3001 gggctaaagg tcgtgcgcct ctgcgccaag agccgtgagg ccatcgactc cccggtgtct
     3061 tttctggccc tgcacaacca gatcaggaac atggacagca tgcctgagct gcagaagctg
     3121 cagcagctga aagacgagac tggggagctg tcgtctgccg acgagaagcg gtaccgggcc
     3181 ttgaagcgca ccgcagagag agagctgctg atgaacgcag atgtcatctg ctgcacatgt
     3241 gtgggcgccg gtgacccgag gctggccaag atgcagttcc gctccatttt aatcgacgaa
     3301 agcacccagg ccaccgagcc ggagtgcatg gttcccgtgg tcctcggggc caagcagctg
     3361 atccttgtag gcgaccactg ccagctgggc ccagtggtga tgtgcaagaa ggcggccaag
     3421 gccgggctgt cacagtcgct cttcgagcgc ctggtggtgc tgggcatccg gcccatccgc
     3481 ctgcaggtcc agtaccggat gcaccctgca ctcagcgcct tcccatccaa catcttctac
     3541 gagggctccc tccagaatgg tgtcactgca gcggatcgtg tgaagaaggg atttgacttc
     3601 cagtggcccc aacccgataa accgatgttc ttctacgtga cccagggcca agaggagatt
     3661 gccagctcgg gcacctccta cctgaacagg accgaggctg cgaacgtgga gaagatcacc
     3721 acgaagttgc tgaaggcagg cgccaagccg gaccagattg gcatcatcac gccctacgag
     3781 ggccagcgct cctacctggt gcagtacatg cagttcagcg gctccctgca caccaagctc
     3841 taccaggagg tggagatcgc cagtgtggac gcctttcagg gacgcgagaa ggacttcatc
     3901 atcctgtcct gtgtgcgggc caacgagcac caaggcattg gctttttaaa tgaccccagg
     3961 cgtctgaacg tggccctgac cagagcaagg tatggcgtca tcattgtggg caacccgaag
     4021 gcactatcaa agcagccgct ctggaaccac ctgctgaact actataagga gcagaaggtg
     4081 ctggtggagg ggccgctcaa caacctgcgt gagagcctca tgcagttcag caagccacgg
     4141 aagctggtca acactatcaa cccgggagcc cgcttcatga ccacagccat gtatgatgcc
     4201 cgggaggcca tcatcccagg ctccgtctat gatcggagca gccagggccg gccttccagc
     4261 atgtacttcc agacccatga ccagattggc atgatcagtg ccggccctag ccacgtggct
     4321 gccatgaaca ttcccatccc cttcaacctg gtcatgccac ccatgccacc gcctggctat
     4381 tttggacaag ccaacgggcc tgctgcaggg cgaggcaccc cgaaaggcaa gactggtcgt
     4441 gggggacgcc agaagaaccg ctttgggctt cctggaccca gccagactaa cctccccaac
     4501 agccaagcca gccaggatgt ggcgtcacag cccttctctc agggcgccct gacgcagggc
     4561 tacatctcca tgagccagcc ttcccagatg agccagcccg gcctctccca gccggagctg
     4621 tcccaggaca gttaccttgg tgacgagttt aaatcacaaa tcgacgtggc gctctcacag
     4681 gactccacgt accagggaga gcgggcttac cagcatggcg gggtgacggg gctgtcccag
     4741 tattaaaagg tggcggcgga agagctaagc aacgtggctt agtccatcag catcttattc
     4801 tgggtaataa aaaaaaaaaa aaagggcggc cgctctagag gatccaccgg atctagataa
     4861 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4921 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4981 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     5101 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5161 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5221 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5281 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5341 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5401 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5461 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5521 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     5581 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5641 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     5701 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5761 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5881 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5941 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6001 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     6061 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     6121 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     6181 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     6241 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     6301 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     6361 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     6421 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6481 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6541 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6601 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6661 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6721 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6781 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6841 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6901 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6961 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     7021 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     7081 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     7141 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     7201 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     7261 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     7321 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7381 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7621 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8101 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8161 aaccgtatta ccgccatgca t
//
