LOCUS       sequence-45439-e        7278 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7278)
  AUTHORS   Azpiazu I, Gautam N
  TITLE     A fluorescence resonance energy transfer-based sensor indicates that
            receptor access to a G protein is unrestricted in a living mammalian
            cell.
  JOURNAL   J Biol Chem. 2004 Jun 25;279(26):27709-18. Epub 2004 Apr 12.
  PUBMED    15078878
REFERENCE   2  (bases 1 to 7278)
  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: "J Biol 
            Chem."; date: "2004-06-25"; volume: "279(26)"; pages: "27709-18. 
            Epub 2004 Apr 12"
COMMENT     Sequence Label: alpha 0-CFP in pdest12.2
FEATURES             Location/Qualifiers
     source          1..7278
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     519..543
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     648..664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     648..664
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        687..705
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     687..704
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    730..854
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(879..884,885..902,903..909)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             963..1622
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1029..1048)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1964..2269
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2148..2167
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2310..2434)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2465..2484)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2466..2484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2495..2512)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2495..2511)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2504..2526)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2916..3050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2953..2972)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3007..3026
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3176..3631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3263..3282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3473..3494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3750..4107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3750..3941
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3773..3793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3958..4093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4158..4952
                     /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(4212..4231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4822..4841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5016..5064
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5105..5124)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5224..5246
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5284..5302)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5370..5474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5475..5543,5544..6335)
                     /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     complement(5693..5712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6506..7094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6995..7014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7248..7265
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcgcgaatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggga cggataacaa tttcacacag gaaacagcta
      661 tgaccattag gcctttgcaa aaagctattt aggtgacact atagaaggta cgcctgcagg
      721 taccggatca caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
      781 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
      841 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
      901 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
      961 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1021 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1081 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     1141 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     1201 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     1261 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     1321 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     1381 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     1441 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     1501 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc
     1561 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     1621 aaacgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     1681 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtgtgc
     1741 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     1801 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     1861 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     1921 gaaatgaacg gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta
     1981 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     2041 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     2101 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     2161 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     2221 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     2281 ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     2341 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     2401 acgtttctcg ttcagctttc ttgtacaaag tggtgatcgc gtgcatgcga cgtcatagct
     2461 ctctccctat agtgagtcgt attataagct aggcactggc cgtcgtttta caacgtcgtg
     2521 actgggaaaa ctgctagctt gggatctttg tgaaggaacc ttacttctgt ggtgtgacat
     2581 aattggacaa actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg
     2641 tataatgtgt taaactagct gcatatgctt gctgcttgag agttttgctt actgagtatg
     2701 atttatgaaa atattataca caggagctag tgattctaat tgtttgtgta ttttagattc
     2761 acagtcccaa ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca
     2821 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2881 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2941 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3001 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gatcctgcat
     3061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggctggcg taatagcgaa
     3121 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     3181 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3241 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3301 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3361 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3421 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3481 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3541 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaat atttaacgcg
     3601 aattttaaca aaatattaac gtttacaatt tcgcctgatg cggtattttc tccttacgca
     3661 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     3721 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3781 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3841 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4021 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4081 ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac
     4141 ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4201 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4261 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4321 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4381 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4441 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4501 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4561 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4621 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4681 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4741 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4801 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4861 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4921 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4981 ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac
     5041 atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatccgcgta tggtgcactc
     5101 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5161 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5221 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5281 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5341 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5401 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5461 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5581 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5641 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5701 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5761 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5881 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5941 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6001 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6061 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6181 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6301 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     6901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7141 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7201 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7261 gcgaggaagc ggaagagc
//
