LOCUS       sequence-247746-        8191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express 3xFLAG tagged DAP5 protein from GUG start codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8191)
  AUTHORS   Kearse MG, Goldman DH, Choi J, Nwaezeapu C, Liang D, Green KM, 
            Goldstrohm AC, Todd PK, Green R, Wilusz JE
  TITLE     Ribosome queuing enables non-AUG translation to be resistant to 
            multiple protein synthesis inhibitors.
  JOURNAL   Genes Dev. 2019 Jun 6. pii: gad.324715.119. doi: 
            10.1101/gad.324715.119.
  PUBMED    31171704
REFERENCE   2  (bases 1 to 8191)
  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: "Genes Dev. 
            2019 Jun 6. pii: gad.324715.119. doi: 10.1101/gad.324715.119."
COMMENT     Sequence Label: pcDNA3.1(+)/GUG-DAP5-3XFLAG
FEATURES             Location/Qualifiers
     source          1..8191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..87
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(306..410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(485..504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        676..1055
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1056..1259
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1210..1230
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1304..1323
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1304..1322
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4111..4176
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4223..4240)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4229..4453
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4499..4927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4586..4605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4796..4817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4936..4956)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4941..5270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5121..5256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5183..5202
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5337..6131
                     /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(5391..5410)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6001..6020
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6305..6426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6342..6361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6396..6415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6475..6491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6475..6491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6488..6510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6499..6515
                     /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(6523..6529,6530..6547,6548..6553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6568..6589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6706..6723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6877..7465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6957..6976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7636..8191,1..236,237..305))
                     /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"
ORIGIN
        1 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
       61 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
      121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
      181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
      241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
      301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      421 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
      481 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
      541 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
      601 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      661 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      721 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      781 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      841 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      901 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      961 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1021 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1081 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1141 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     1201 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     1261 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     1321 gggagaccca agctggctag cgtttaaact taagcttggc aatccggtac tgttggtaaa
     1381 taagccaccg tggagagtgc gattgcagaa gggggtgctt ctcgtttcag tgcttcttcg
     1441 ggcggaggag gaagtagggg tgcacctcag cactatccca agactgctgg caacagcgag
     1501 ttcctgggga aaaccccagg gcaaaacgct cagaaatgga ttcctgcacg aagcactaga
     1561 cgagatgaca actccgcagc aaacaactcc gcaaacgaaa aagaacgaca tgatgcaatc
     1621 ttcaggaaag taagaggcat actaaataag cttactcctg aaaagtttga caagctatgc
     1681 cttgagctcc tcaatgtggg tgtagagtct aaactcatcc ttaaaggggt catactgctg
     1741 attgtggaca aagccctaga agagccaaag tatagctcac tgtatgctca gctatgtctg
     1801 cgattggcag aagatgcacc aaactttgat ggcccagcag cagagggtca accaggacag
     1861 aagcaaagca ccacattcag acgcctccta atttccaaat tacaagatga atttgaaaac
     1921 cgaactagaa atgttgatgt ctatgataag cgtgaaaatc ccctcctccc cgaggaggag
     1981 gaacagagag ccattgctaa gatcaagatg ttgggaaaca tcaaattcat tggagagctt
     2041 ggcaagcttg atcttattca cgaatctatc cttcataagt gcatcaaaac acttttggaa
     2101 aagaagaaga gagtccaact caaagatatg ggagaggatt tggagtgcct ctgtcagata
     2161 atgaggacag tgggacctag attagaccat gaacgagcca agtccttaat ggatcagtac
     2221 tttgcccgaa tgtgctcctt gatgttaagt aaggaattgc cagcaaggat tcgtttcctg
     2281 ctgcaggata ccgtagagtt gcgagaacac cattgggttc ctcgcaaggc ttttcttgac
     2341 aatggaccaa agacgatcaa tcaaattcgt caagatgcag taaaagatct aggggtgttt
     2401 attcctgctc ctatggctca agggatgaga agtgacttct ttctggaggg accgttcatg
     2461 ccacccagga tgaaaatgga tagggaccca cttggaggac ttgctgatat gtttggacaa
     2521 atgccaggta gcggaattgg tactggtcca ggagttatcc aggatagatt ttcacccacc
     2581 atgggacgtc atcgttcaaa tcaactcttc aatggccatg ggggacacat catgcctccc
     2641 acacaatcgc agtttggaga gatgggaggc aagtttatga aaagccaggg gctaagccag
     2701 ctctaccata accagagtca gggactctta tcccagctgc aaggacagtc gaaggatatg
     2761 ccacctcggt tttctaagaa aggacagctt aatgcagatg agattagcct gaggcctgct
     2821 cagtcgttcc taatgaataa aaatcaagtg ccaaagcttc agccccagat aactatgatt
     2881 cctcctagtg cacaaccacc acgcactcaa acaccacctc tgggacagac acctcagctt
     2941 ggtctcaaaa ctaatccacc acttatccag gaaaagcctg ccaagaccag caaaaagcca
     3001 ccaccgtcaa aggaagaact ccttaaacta actgaaactg ttgtgactga atatctaaat
     3061 agtggaaatg caaatgaggc tgtcaatggt gtaagagaaa tgagggctcc taaacacttt
     3121 cttcctgaga tgttaagcaa agtaatcatc ctgtcactag atagaagcga tgaagataaa
     3181 gaaaaagcaa gttctttgat cagtttactc aaacaggaag ggatagccac aagtgacaac
     3241 ttcatgcagg ctttcctgaa tgtattggac cagtgtccca aactggaggt tgacatccct
     3301 ttggtgaaat cctatttagc acagtttgca gctcgtgcca tcatttcaga gctggtgagc
     3361 atttcagaac tagctcaacc actagaaagt ggcacccatt ttcctctctt cctactttgt
     3421 cttcagcagt tagctaaatt acaagatcga gaatggttaa cagaactttt tcaacaaagc
     3481 aaggtcaata tgcagaaaat gctcccagaa attgatcaga ataaggaccg catgttggag
     3541 attttggaag gaaagggact gagtttctta ttcccactcc tcaaattgga gaaggaactg
     3601 ttgaagcaaa taaagttgga tccatctcct caaaccatat ataaatggat taaagataac
     3661 atctctccca aacttcatgt agataaagga tttgtgaaca tcttaatgac tagcttctta
     3721 cagtacattt ctagtgaagt aaaccccccc agcgatgaaa cagattcatc ctctgctcct
     3781 tccaaagaac agttagagca ggaaaaacaa ctactactat ctttcaagcc agtaatgcag
     3841 aaatttcttc atgatcacgt tgatctacaa gtcagtgccc tgtatgctct ccaggtgcac
     3901 tgctataaca gcaacttccc aaaaggcatg ttacttcgct tttttgtgca cttctatgac
     3961 atggaaatta ttgaagaaga agctttcttg gcttggaaag aagatataac ccaagagttt
     4021 ccgggaaaag gcaaggcttt gttccaggtg aatcagtggc taacctggtt agaaactgct
     4081 gaagaagaag aatcagagga agaagctgac gactacaaag accatgacgg tgattataaa
     4141 gatcatgaca tcgattacaa ggatgacgat gacaagtaat aaggccgcga ctctagaggg
     4201 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4261 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4321 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4381 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4441 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4921 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5041 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5101 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5161 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5221 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5281 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5341 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5401 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5461 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5521 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5581 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5641 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5701 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5761 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5821 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5881 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5941 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6001 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6061 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6121 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6181 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6241 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6301 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6361 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6421 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8161 cgtaagatgc ttttctgtga ctggtgagta c
//
