LOCUS       sequence-252676-        5711 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 5711)
  AUTHORS   Watson MJ, Worthington EN, Clunes LA, Rasmussen JE, Jones L, Tarran 
            R
  TITLE     Defective adenosine-stimulated cAMP production in cystic fibrosis 
            airway epithelia: a novel role for CFTR in cell signaling.
  JOURNAL   FASEB J. 2011 Sep;25(9):2996-3003. Epub 2011 May 31.
  PUBMED    21628448
REFERENCE   2  (bases 1 to 5711)
  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: "FASEB J."; 
            date: "2011-09"; volume: "25(9)"; pages: "2996-3003. Epub 2011 May 
            31"
COMMENT     Sequence Label: pEYFP-N1-A2BR
FEATURES             Location/Qualifiers
     source          1..5711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1287..1296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1293..1295,1296..1298,1299..2012)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1338..1359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1599..1618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1946..1967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2135..2256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2172..2191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2226..2245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2263..2718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2400..2421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2612..2631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2745..2849
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2851..3208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2851..3042
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2874..2894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3059..3194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3243..4037
                     /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             3243..4037
                     /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(3297..3316)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3907..3926
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4225..4244)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4269..4316
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4645..5233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5134..5153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5408..5711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc ccaccatgct gctggagaca caggacgcgc
      301 tgtacgtggc gctggagctg gtcatcgccg cgctttcggt ggcgggcaac gtgctggtgt
      361 gcgccgcggt gggcacggcg aacactctgc agacgcccac caactacttc ctggtgtccc
      421 tggctgcggc cgacgtggcc gtggggctct tcgccatccc ctttgccatc accatcagcc
      481 tgggcttctg cactgacttc tacggctgcc tcttcctcgc ctgcttcgtg ctggtgctca
      541 cgcagagctc catcttcagc cttctggccg tggcagtcga cagatacctg gccatctgtg
      601 tcccgctcag gtataaaagt ttggtcacgg ggacccgagc aagaggggtc attgctgtcc
      661 tctgggtcct tgcctttggc atcggattga ctccattcct ggggtggaac agtaaagaca
      721 gtgccaccaa caactgcaca gaaccctggg atggaaccac gaatgaaagc tgctgccttg
      781 tgaagtgtct ctttgagaat gtggtcccca tgagctacat ggtatatttc aatttctttg
      841 ggtgtgttct gcccccactg cttataatgc tggtgatcta cattaagatc ttcctggtgg
      901 cctgcaggca gcttcagcgc actgagctga tggaccactc gaggaccacc ctccagcggg
      961 agatccatgc agccaagtca ctggccatga ttgtggggat ttttgccctg tgctggttac
     1021 ctgtgcatgc tgttaactgt gtcactcttt tccagccagc tcagggtaaa aataagccca
     1081 agtgggcaat gaatatggcc attcttctgt cacatgccaa ttcagttgtc aatcccattg
     1141 tctatgctta ccggaaccga gacttccgct acacttttca caaaattatc tccaggtatc
     1201 ttctctgcca agcagatgtc aagagtggga atggtcaggc tggggtacag cctgctctcg
     1261 gtgtgggcct aggggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1321 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1381 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1441 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc
     1501 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1561 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1621 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1681 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1741 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1801 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1861 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     1921 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1981 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     2041 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2101 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2161 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2221 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2281 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2341 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2401 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2461 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2521 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2581 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2641 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2701 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2761 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2821 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2881 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2941 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3001 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3061 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3121 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3181 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3241 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3301 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3361 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3421 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3481 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3541 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3601 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3661 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3721 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3781 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3841 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3901 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3961 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4021 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4081 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4141 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4201 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4261 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4321 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4381 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4441 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5341 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5401 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5461 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5521 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5581 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     5641 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     5701 ctattaccat g
//
