LOCUS       sequence-99587-e        7592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of EGFP-tagged human CUGBP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7592)
  TITLE     Charlet-Berguerand Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7592)
  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            "Unpublished"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: pDest eGFP CUGBP1
FEATURES             Location/Qualifiers
     source          1..7592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      857..866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(863..865,866..868,869..1579)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(863..865,866..868,869..1579)
                     /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(908..929)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1169..1188)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1516..1537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1625..1649
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3126..3150)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3183..3200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3189..3413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3459..3887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3546..3565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3756..3777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3896..3916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3901..4230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4081..4216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4143..4162
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4297..5091
                     /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(4351..4370)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4961..4980
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5265..5386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5302..5321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5356..5375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5435..5451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5435..5451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5448..5470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5459..5475
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5483..5489,5490..5507,5508..5513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5528..5549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5666..5683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5837..6425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5917..5936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6596..7387,7388..7456))
                     /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     7219..7238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7457..7561)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1561 catggacgag ctgtacaagt actcagatct cgagctctac aaaagcggtt ccggaccgga
     1621 tcaaacaagt ttgtacaaaa aagcaggctc cgaattcgcc cttatgaacg gcaccctgga
     1681 ccacccagac caaccagatc ttgatgctat caagatgttt gtgggccagg ttccaaggac
     1741 ctggtctgaa aaggacttgc gggaactctt cgaacagtat ggtgctgtgt atgaaatcaa
     1801 cgtcctaagg gataggagcc aaaacccgcc tcagagcaaa gggtgctgtt ttgttacatt
     1861 ttacacccgt aaagctgcat tagaagctca gaatgctctt cacaacatga aagtcctccc
     1921 agggatgcat caccctatac agatgaaacc tgctgacagt gagaagaaca atgcagtgga
     1981 agacaggaag ctgtttattg gtatgatttc caagaagtgc actgaaaatg acatccgagt
     2041 catgttctct tcgtttggac agattgaaga atgccggata ttgcggggac ctgatggcct
     2101 gagccgaggt tgtgcatttg tgacttttac aacaagagcc atggcacaga cggctatcaa
     2161 ggcaatgcac caagcacaga ccatggaggg ttgctcatca cccatggtgg taaaatttgc
     2221 tgatacacag aaggacaaag aacagaagag aatggcccag cagctccagc agcagatgca
     2281 gcaaatcagc gcagcatctg tgtggggaaa ccttgctggt ctaaatactc ttggacccca
     2341 gtatttagca ctccttcagc agactgcctc ctctgggaac ctcaacaccc tgagcagcct
     2401 ccacccaatg ggagggttga atgcaatgca gttacagaat ttggctgcac tagctgctgc
     2461 agctagtgca gctcagaaca caccaagtgg taccaatgct ctcactacat ccagcagtcc
     2521 cctcagcgtg ctcactagtt cagggtcctc acctagctct agcagcagta attctgtcaa
     2581 ccccatagcc tcacttggag ccctgcagac attagctgga gcaacggctg gcctcaatgt
     2641 tggctctttg gcaggaatgg ctgctttaaa tggtggcctg ggcagcagtg gcctttccaa
     2701 tggcaccggg agcaccatgg aggccctcac tcaggcctac tcgggtatcc agcaatatgc
     2761 tgctgctgcg ctccccactc tgtacaacca gaatcttctg acacagcaga gtattggtgc
     2821 tgctggaagc cagaaggaag gtccagaggg agccaacctg ttcatctacc acctgcccca
     2881 ggagtttggt gatcaggacc tgctgcagat gtttatgccc tttgggaatg tcgtgtctgc
     2941 caaggttttc atagacaagc agacaaacct gagcaagtgt tttggttttg taagttacga
     3001 caatcctgtt tcggcccaag ctgccatcca gtccatgaac ggctttcaga ttggcatgaa
     3061 gcggcttaaa gtgcagctca aacgttcgaa gaatgacagc aagccctact gaaagggcga
     3121 attcgaccca gctttcttgt acaaagtggt ggataattaa ttaagataaa cccgctgatc
     3181 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3241 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3301 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3361 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3421 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3481 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3541 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3601 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3661 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3721 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3781 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3841 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3901 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4021 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4081 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4141 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4201 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4261 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4321 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4381 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4441 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4501 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4561 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4621 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4681 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4741 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4801 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4861 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4921 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4981 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5041 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5101 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5161 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5221 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5281 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5341 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5401 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5461 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5521 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5581 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5641 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5701 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5761 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5821 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5881 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5941 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6001 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6061 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6121 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6181 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6241 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6301 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6361 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6421 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6481 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6541 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6601 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6661 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6721 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6781 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6841 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6901 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6961 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7021 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7081 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7141 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7201 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7261 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7321 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7381 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7441 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7501 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7561 gcacatttcc ccgaaaagtg ccacctgacg tc
//
