LOCUS       sequence-488138-        7401 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_115, derived from human fusion protein NDRG1_ERG; TMPRSS2_ERG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7401)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_115
FEATURES             Location/Qualifiers
     source          1..7401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(934..1644,1645..1647,1648..1650))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(976..997)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1325..1344
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1584..1605
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1647..1656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1732..1935)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1761..1781)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1936..2239
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2496..2540)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2725..2958
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    3318..3435
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3927..4052
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4099..4356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4358..4737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4768..4872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4873..4941,4942..5733)
                     /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(5091..5110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5904..6492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6393..6412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6646..6663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6731..6865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6755..6774)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6809..6828
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6871..7066)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6885..7020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7125..7146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7161..7166,7167..7184,7185..7191)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7199..7215
                     /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)."
     primer_bind     7204..7226
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7223..7239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7223..7239
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctgctagctc gagcagctga agctttattg aggcttaagc
      361 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
      421 ccagtacagg caaaaagcag atcttgtctt ctttgggagt gaattagccc ttccagtccc
      481 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
      541 ggtaccgtaa tcttatctag aaccggttca gtctgcgggc acgatcacac gacgttcgtt
      601 agttgtcata tttggcggag gcatgtgttt ttcctccata taagaaccgt agttcatacc
      661 caccgtatcg ggtgatccta ccatcttgcc gcccttggcc acagaacatt cgtcggggct
      721 gttgcgggac ccgtttacct gggacggatt gcactccatc ttaatcgtga cgcgggctgg
      781 cggttgcgac aaccagtctt gttgcggtac acggggtgac attttcgatg tctgaccgta
      841 atcggaagag gaggatgcag tcatctcagt cttggccaaa tgtggagttc cccgggaagc
      901 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
      961 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1021 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1081 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1141 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1201 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1261 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1321 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     1381 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     1441 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1501 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     1561 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1621 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     1681 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     1741 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     1801 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     1861 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     1921 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     1981 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2041 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2101 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2161 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2221 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2281 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     2341 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     2401 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     2461 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     2521 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     2581 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     2641 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     2701 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     2761 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     2821 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     2881 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     2941 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     3001 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     3061 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     3121 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     3181 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     3241 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     3301 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     3361 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     3421 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     3481 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     3541 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     3601 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     3661 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     3721 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     3781 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     3841 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     3901 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     3961 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     4021 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     4081 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     4141 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     4201 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     4261 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     4321 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     4381 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4441 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4501 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4561 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4621 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4681 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     4741 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4861 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4921 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4981 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5041 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5101 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5161 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5221 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5281 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5341 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5401 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5461 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5521 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5581 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5641 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5701 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5761 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5821 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5881 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5941 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6001 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6061 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6121 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6181 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6241 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6301 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6361 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6421 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6481 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6541 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6601 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6661 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6721 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6781 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6841 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     6901 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     6961 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7021 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     7081 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7141 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7201 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     7261 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     7321 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     7381 aaataacaca aaacaaaata t
//
