LOCUS       sequence-488137-        7329 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_114, 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 7329)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7329)
  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_114
FEATURES             Location/Qualifiers
     source          1..7329
                     /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(862..1572,1573..1575,1576..1578))
                     /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(904..925)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1253..1272
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1512..1533
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1575..1584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1660..1863)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1689..1709)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1864..2167
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2424..2468)
                     /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    2653..2886
                     /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    3246..3363
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3855..3980
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4027..4284
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4286..4665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4696..4800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4801..4869,4870..5661)
                     /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(5019..5038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5832..6420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6321..6340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6574..6591
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6659..6793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6683..6702)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6737..6756
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6799..6994)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6813..6948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7053..7074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7089..7094,7095..7112,7113..7119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7127..7143
                     /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     7132..7154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7151..7167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7151..7167
                     /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 atagtaggtg cccaagtgag acggcatgtg
      601 actggttggc aagcgcgtgt tagggtagat gccccccgtg ggtgagttcc aatatggatt
      661 aggtgcagca aagaagctag agctagtcac tggtaaagcc gggggatgcg gtgccacaaa
      721 gttcatcttc tgtggatgag cgtgatacga tcccatgtac ggcaagtctg atggatattt
      781 ataaagcgaa ctctctggag gatgtggttg taacgcttgc cgggaagcgg ccgcacttcc
      841 cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
      901 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
      961 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1021 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     1081 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     1141 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1201 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1261 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1321 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     1381 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     1441 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1501 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1561 ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg
     1621 cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta tatagacctc
     1681 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     1741 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     1801 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     1861 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     1921 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     1981 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     2041 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     2101 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     2161 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     2221 ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag gtgggtctga
     2281 aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact
     2341 attcttaaac ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa
     2401 tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt
     2461 tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa
     2521 ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat
     2581 ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa
     2641 ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc
     2701 tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc
     2761 ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc
     2821 agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa
     2881 gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt
     2941 gctactccta atggttcaat ttttactact ttatatttat ataattcact tctccaattg
     3001 tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta atcctcatcc
     3061 tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct aatgatcttt
     3121 gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac tgccccttca
     3181 cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc cctgtaataa
     3241 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     3301 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     3361 aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc ctgatctctt
     3421 acctgtccta taattttctt taattcttta ttcatagatt ctactactcc ttgactttgg
     3481 ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt tttcttactt
     3541 ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt
     3601 tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct
     3661 gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac
     3721 aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat
     3781 ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc
     3841 ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa
     3901 atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc
     3961 cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg
     4021 cgccactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     4081 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     4141 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     4201 gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc tcgccactcc
     4261 ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg tagatgtact
     4321 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     4381 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     4441 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     4501 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     4561 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     4621 accccgtaat tgattactat taataactag tcaataatca atgtcgaccc aggtggcact
     4681 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4741 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4861 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4921 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4981 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5041 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5101 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5161 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5221 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5281 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5341 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5401 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5461 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5521 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5581 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5641 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5701 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5761 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5821 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5881 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5941 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6001 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6061 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6121 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6181 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6241 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6301 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6361 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6421 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6481 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6541 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6601 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcgga
     6661 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     6721 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     6781 caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     6841 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     6901 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     6961 gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga caggtttccc
     7021 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7081 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7141 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct gcaggtcgag
     7201 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
     7261 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
     7321 acaaaatat
//
