LOCUS       sequence-160813-        6782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Avi-tagged RPL1 and humanized birA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6782)
  AUTHORS   Ingolia NT, Brar GA, Stern-Ginossar N, Harris MS, Talhouarne GJ, 
            Jackson SE, Wills MR, Weissman JS
  TITLE     Ribosome profiling reveals pervasive translation outside of 
            annotated protein-coding genes.
  JOURNAL   Cell Rep. 2014 Sep 11;8(5):1365-79. doi: 
            10.1016/j.celrep.2014.07.045. Epub 2014 Aug 21.
  PUBMED    25159147
REFERENCE   2  (bases 1 to 6782)
  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"; doi: 
            "10.1016/j.celrep.2014.07.045"; journalName: "Cell Rep"; date: 
            "2014-09-11- 11"; volume: "8"; issue: "5"; pages: "1365-79"
COMMENT     Sequence Label: pNTI194
FEATURES             Location/Qualifiers
     source          1..6782
                     /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"
     CDS             1577..1597
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1595..1639
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1649..1705
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2669..2692
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2734..2751)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2740..2964
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3097..3116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3248..3295
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3303..4325
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3303..4325
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4455..4576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4492..4511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4546..4565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4625..4641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4625..4641)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4638..4660)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4649..4665
                     /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(4673..4679,4680..4697,4698..4703))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4718..4739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4856..4873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5027..5615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5107..5126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5786..6577,6578..6646))
                     /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     6409..6428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6647..6751)
                     /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 ccatgagcag caaagtctct cgcgacaccc tgtacgaggc
      901 ggtgcgggaa gtcctgcacg ggaaccagcg caagcgccgc aagttcctgg agacggtgga
      961 gttgcagatc agcttgaaga actatgatcc ccagaaggac aagcgcttct cgggcaccgt
     1021 caggcttaag tccactcccc gccctaagtt ctctgtgtgt gtcctggggg accagcagca
     1081 ctgtgacgag gctaaggccg tggatatccc ccacatggac atcgaggcgc tgaaaaaact
     1141 caacaagaat aaaaaactgg tcaagaagct ggccaagaag tatgatgcgt ttttggcctc
     1201 agagtctctg atcaagcaga ttccacgaat cctcggccca ggtttaaata aggcaggaaa
     1261 gttcccttcc ctgctcacac acaacgaaaa catggtggcc aaagtggatg aggtgaagtc
     1321 cacaatcaag ttccaaatga agaaggtgtt atgtctggct gtagctgttg gtcacgtgaa
     1381 gatgacagac gatgagcttg tgtataacat tcacctggct gtcaacttct tggtgtcatt
     1441 gctcaagaaa aactggcaga atgtccgggc cttatatatc aagagcacca tgggcaagcc
     1501 ccagcgccta tatggaggtt cttcaggaag tgggagttca ggaggctccg gatcaagcgg
     1561 cagtagtggt ggaagcgaaa acctgtactt ccagggcctg aatgacatct ttgaggccca
     1621 gaagattgaa tggcacgagg gaagcggagc tactaacttc agcctgctga agcaggctgg
     1681 agacgtggag gagaaccctg gacctatgaa ggacaacacc gtgcccctga agctgatcgc
     1741 cctgctggcc aatggcgagt tccactctgg cgagcagctg ggcgaaaccc tgggaatgtc
     1801 tgccgaggcc atcaacaagc acatccagac cctgcgggat tggggcgtgg acgtgttcac
     1861 agtgcctggc aagggctaca gcctgcccga gcctatccag ctgctgaacg ccgagaagat
     1921 cctgagccag ctggacgatg gcagcgtggc agtgctgccc gtgatcgaca gcaccaacca
     1981 gtacctgctg gaccggatcg gcgagctgaa aagcggagat gcctgcgtgg ccgagtatca
     2041 gcacgccgga agggggagaa gaggccgcaa gtggttcagc cccttcggcg ccaatctgta
     2101 cctgagcatg ttctggcggc tggaacaggg acctgccgcc gctattggac tgagcctcgt
     2161 gatcggcatc gtgatggccg aggtgctgag aaagctgggc gccgacaaag tgcgcgtgaa
     2221 gtggcccaac gacctgtatc tgcaggaccg gaagctggcc ggcatcctgg tggaactgac
     2281 aggcaaaaca ggcgacgccg cccagatcgt gattggagcc ggcatcaata tggccatgcg
     2341 gcgggtggaa gagtccgtcg tgaatcaggg ctggatcacc ctgcaggaag ccggaatcaa
     2401 cctggacaga aacaccctgg ccgccatgct gatcagagag ctgagagccg ccctggaact
     2461 gttcgagcag gaaggcctga ccccctacct gtccagatgg gagaagctgg acaacttcat
     2521 caacagaccc gtgaaactga tcatcggcga caaagagatc ttcggcatca gccggggcat
     2581 cgacaagcag ggcgctctgc tgctggaaca ggacggcatc atcaagccct ggatgggcgg
     2641 cgagatcagc ctgagatctg ccgagaagga ctacaaggac gatgacgata agtgagctcg
     2701 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2761 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2821 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2881 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2941 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3001 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3061 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3121 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3181 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3241 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     3301 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     3361 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     3421 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     3481 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     3541 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     3601 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     3661 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     3721 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     3781 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     3841 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     3901 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     3961 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     4021 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     4081 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     4141 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     4201 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     4261 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     4321 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4381 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4441 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4501 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4561 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4621 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4681 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4741 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4801 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4861 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4921 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4981 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5041 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5101 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5161 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5221 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5281 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5341 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5401 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5461 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5521 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5581 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5641 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5701 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5761 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5821 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5881 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5941 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6001 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6061 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6121 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6181 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6241 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6301 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6361 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6421 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6481 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6541 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6601 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6661 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6721 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6781 tc
//
