LOCUS       sequence-488136-        7290 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_113, 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 7290)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7290)
  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_113
FEATURES             Location/Qualifiers
     source          1..7290
                     /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(823..1533,1534..1536,1537..1539))
                     /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(865..886)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1214..1233
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1473..1494
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1536..1545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1621..1824)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1650..1670)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1825..2128
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2385..2429)
                     /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    2614..2847
                     /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    3207..3324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3816..3941
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3988..4245
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4247..4626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4762..4830,4831..5622)
                     /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(4980..4999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5793..6381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6282..6301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6535..6552
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6620..6754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6644..6663)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6698..6717
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6760..6955)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6774..6909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7014..7035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7050..7055,7056..7073,7074..7080)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7088..7104
                     /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     7093..7115
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7112..7128
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7112..7128
                     /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 cggatttgct aaacgggagc ttgtcggacc
      601 caggatctga taggggtcca gttgtggacg ttgatcctct gtcttgggta ccgttgatgg
      661 actgggctgg gcagccttac tttggggagt gggatgaccg tgtccggtcc acgcggagcg
      721 gcggggcggt tcgtagggca aatcgggacg agttgtgatg cgctgtgttg cctcagggta
      781 ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca gctcgtccat
      841 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      901 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
      961 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     1021 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     1081 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     1141 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1201 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     1261 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1321 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     1381 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     1441 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1501 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg
     1561 ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt tcactaaacg
     1621 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     1681 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     1741 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     1801 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     1861 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     1921 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     1981 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     2041 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     2101 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     2161 ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct cggtcccctc
     2221 ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac tctattcact
     2281 atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat cattatgaat
     2341 aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact tgcccattta
     2401 tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc ttcaattaag
     2461 gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca ggtcgtgtga
     2521 ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac agcagtggtg
     2581 caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt taggtatctt
     2641 tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag ttgcaacaga
     2701 tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat accagacaat
     2761 aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct
     2821 cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct ctgcaccact
     2881 cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac tttatattta
     2941 tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt actaaacttt
     3001 tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc catctgtttt
     3061 ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc actattatct
     3121 tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt ccaaagtgga
     3181 tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat
     3241 tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc
     3301 ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac tgctgtctta
     3361 agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt attcatagat
     3421 tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga tcctgctgct
     3481 tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     3541 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     3601 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     3661 cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag ctccctgctt
     3721 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     3781 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     3841 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     3901 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     3961 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     4021 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     4081 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     4141 ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc ttatatgcag
     4201 gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg gtaatagcga
     4261 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4321 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4381 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4441 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4501 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4561 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     4621 aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4801 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4861 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4921 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4981 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5041 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5101 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5161 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5221 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5281 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5341 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5401 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5461 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5521 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5581 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6421 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6481 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6541 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6601 ccgattcatt aatgcagcgg atccagacat gataagatac attgatgagt ttggacaaac
     6661 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     6721 atttgtaacc attataagct gcaataaaca agttagcttt ttgcaaaagc ctaggcctcc
     6781 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     6841 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     6901 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcggatc
     6961 cgctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7021 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7081 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7141 aagcttgggc tgcaggtcga gggatctcca taagagaaga gggacagcta tgactgggag
     7201 tagtcaggag aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt
     7261 taaagaaaaa aataacacaa aacaaaatat
//
