LOCUS       sequence-359479-        8023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orf 73 tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8023)
  TITLE     Dittmer lab plasmids Jan 2022
REFERENCE   2  (bases 1 to 8023)
  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"
COMMENT     Sequence Label: pOME0349
FEATURES             Location/Qualifiers
     source          1..8023
                     /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     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    914..938
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      977..986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2750..2815
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2836..2860)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2871..2873,2874..2876,2877..3590)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2904..2932)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3527..3547
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3612..3629)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3618..3842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3888..4316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3975..3994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4185..4206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4325..4345)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4330..4659
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4510..4645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4572..4591
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4726..5520
                     /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(4780..4799)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5390..5409
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5696..5817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5733..5752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5787..5806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5866..5882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5866..5882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5879..5901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5890..5906
                     /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(5914..5920,5921..5938,5939..5944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5959..5980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6097..6114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6268..6856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6348..6367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7027..7818,7819..7887))
                     /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     7650..7669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7888..7992)
                     /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 gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttgat caaacaagtt tgtacaaaaa agcaggctct ttaaaggaac caattcagtc
      961 gactggatcc ggtaccgcca ccatggcgcc cccgggaatg cgcctgaggt cgggacggag
     1021 caccggcgcg cccttaacga gaggaagttg taggaaacga aacaggtctc cggaaagatg
     1081 tgaccttggc gatgacctac atctacaacc gcgaaggaag catgtcgccg actccgtcga
     1141 cggccgggaa tgtggacccc acaccttgcc tataccagga agtcccacag tgttcacatc
     1201 cgggctgcca gcatttgtgt ctagtcctac tttaccggtg gctcccattc cttcacccgc
     1261 tcccgcaaca cctttacctc caccggcact cttacccccc gtaaccacgt cttcctcccc
     1321 aatccctcca tcccatcctg tgtctccggg gaccacggat actcattctc catctcctgc
     1381 attgccaccc acgcagtctc cagagtcttc tcaaaggcca ccgctttcaa gtcctacagg
     1441 aaggccagac tcttcaacac ctatgcgtcc gccaccctcg cagcagacta cacctccaca
     1501 ctcacccacg actcctccac ccgagcctcc ctccaagtcg tcaccagact ctttagctcc
     1561 gtctaccctg cgtagcctga gaaaaagaag gctatcgtcc ccccaaggtc cctctacact
     1621 aaacccaata tgtcagtcgc ccccagtctc tccccctaga tgtgacttcg ccaaccgtag
     1681 tgtgtacccc ccatgggcca cagagtcccc gatctacgtg ggatcatcca gcgatggcga
     1741 tactccgcca cgccaaccgc ctacatctcc catctccata ggatcatcat ccccgtctga
     1801 gggatcctgg ggtgatgaca cagccatgtt ggtgctcctt gcggagattg cagaagaagc
     1861 atccaagaat gaaaaagaat gttccgaaaa taatcaggct ggcgaggata atggggacaa
     1921 cgagattagc aaggaaagtc aggttgacaa ggatgacaat gacaataagg atgatgagga
     1981 ggagcaggag acagacgagg aggacggggg cgatggaaac aaaacgttga gcatccaaag
     2041 ttcacaagag caggagacgg tggaagagcc cataatcttg cacgggtcgt catccgagga
     2101 cgaaatggaa gtggattacc ctgttgttag cacacatgaa caaattgcca gtagcccacc
     2161 aggagataat acaccagacg atgacccaca acctggccca tctcgcgaat accgctatgt
     2221 actcagaaca tcaccacccc acagacctgg agttcgtatg aggcgcgttc cagttaccca
     2281 cccaaaaaag ccacatccaa gataccaaca accaccggtc ccttacagac agatagatga
     2341 ttgtcctgcg aaagctaggc cacaacacat cttttataga cgctttttgg gaaaggatgg
     2401 aagacgagat ccaaagtgtc aatggaagtt tgcagtgatt ttttggggca atgacccata
     2461 cggacttaaa aaattatctc aggccttcca gtttggagga gtaaaggcag gccccgtgtc
     2521 ctgcttgccc caccctggac cagaccagtc gcccataact tattgtgtat atgtgtattg
     2581 tcagaacaaa gacacaagta agaaagtaca aatggcccgc ctagcctggg aagctagtca
     2641 ccccctggca ggaaacctac aatcttccat agttaagttt aaaaagcccc tgccattaac
     2701 ccagccaggg gaaaaccaag gtcctgggga ctctccacag gaaatgacag actacaaaga
     2761 ccatgacggt gattataaag atcatgatat cgattacaag gatgacgatg acaagtagta
     2821 actcgagata tctagaccca gctttcttgt acaaagtggt tcgatctaga atggctagca
     2881 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     2941 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgctacatac ggaaagctta
     3001 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
     3061 ctttctctta tggtgttcaa tgcttttccc gttatccgga tcatatgaaa cggcatgact
     3121 ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct ttcaaagatg
     3181 acgggaacta caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta
     3241 tcgagttaaa aggtattgat tttaaagaag atggaaacat tctcggacac aaactcgagt
     3301 acaactataa ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
     3361 ctaacttcaa aattcgccac aacattgaag atggatccgt tcaactagca gaccattatc
     3421 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga
     3481 cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt
     3541 ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa tgaattaaac
     3601 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3661 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3721 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3781 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3841 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3901 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3961 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4021 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4081 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4141 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4201 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4261 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4321 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4381 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4501 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4561 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4621 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4681 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4741 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4801 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4861 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4921 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4981 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5041 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5101 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5161 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5221 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5281 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5341 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5401 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5461 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5521 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5701 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5761 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5821 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5881 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5941 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6001 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6061 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6121 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6181 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6241 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6301 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6361 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6421 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6481 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6541 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6601 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6661 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6721 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6781 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6841 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6901 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6961 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7021 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7081 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7141 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7201 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7261 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7321 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7381 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7441 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7501 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7561 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7621 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7681 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7741 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7801 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7861 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
