LOCUS       sequence-488256-        8661 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_027, derived from human protein TAB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8661)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8661)
  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_Vidr_027
FEATURES             Location/Qualifiers
     source          1..8661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3692..4402,4403..4405,4406..4408))
                     /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(3734..3755)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4083..4102
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4342..4363
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4405..4414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4490..4693)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4519..4539)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4694..4997
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5254..5298)
                     /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    5483..5716
                     /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    6076..6193
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6685..6810
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6857..7114
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7116..7495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7526..7630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7631..7699,7700..8491)
                     /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(7849..7868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct gctagctcga gcagctgaag ctttattgag gcttaagcag
     1861 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
     1921 agtacaggca aaaagcagat cttgtcttct ttgggagtga attagccctt ccagtccccc
     1981 cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg
     2041 taccgtaatc ttatctagaa ccggttcaca tttcctcaag actcgggatc tgggaaatag
     2101 aattgctacg ctttaagcgg cgacgtgtca gattgttttc catttcgttc acctcggatt
     2161 taagtttgtc aagttttttc ttctggattt ccagttcacg ctgtaagcgc tccatacgtg
     2221 ctttctgatg aactaaaaga gcttgggtgt acgcggcatc atcagacccc attgacagtt
     2281 tacgcgtctc gctgatgcga tccacatgtg ctaacgtagg atcagtcaaa tgctgaatat
     2341 tttcagtctc gacatagtga tcgggatgat tcagcagatt cgtcaactcg aacgtggggg
     2401 aaaccacacc cggactgacg gcaggcggct tgtttggtga taccgtaatt ttaaacgtat
     2461 atttcgtgtt tggttgggtt acgacaacac gtggagaggt agctgaatta ttgccaatag
     2521 cgcgtgattt tggcgggtga tggtggataa acgccgggcc catagagact tggccgctca
     2581 tgttacggga ggctgctgag gctcctgagt tcgttgagat gaataatgtc ggctgattac
     2641 gcatcacctg ttcatcgccg gtggcggcat tcgccgaaat gtaaacctta ggttgactac
     2701 gactcataag ttcatccgtg ttaggcggcg aggcagcaat gtacactgta ggctgattgc
     2761 ggttaagcgt ctgagaattg acggaagatg aggtagagga ggtgcggggg ccagaagaac
     2821 gcagtttgct agaattattg cgctgaggcg gttccaattt gatctcgatt tgattcttgc
     2881 gaggaccagt cgagatgttt tgaatattgt actgtgagtg cgcactggat tggctgcttc
     2941 cgctggaatg aatagtcggc ggttgacttg tagtcggcga actaatgggc atgtatacgt
     3001 gactggtttg gtgtccttgt tggttaggct gttgcgagga ggtgtgtgac agtgggttag
     3061 aggcaggaca cgtcgtccaa ggtcctggtt ggctaggctg gtatacttgc tgagggttca
     3121 tggggttaaa ctgtgacacc catcccgagt gttgctgggt ctgacgcgtg gtaccgcctg
     3181 gagtagtgat ataagggcga atgtagatgc tgttcccttg aggggagttc aatacgggtg
     3241 gcggcacgcc atgaatatgc aaagaagtag gggtattgcg gccggtttgg atattcgggg
     3301 ccaatgtgac cataatcgga ttgaaacgtg gtgtctgctg actcaaagat gacgtacctt
     3361 ttgaccccaa atggaaaccc aggtgcggtg cagaattgct agcgccggag gaactactca
     3421 tcccaaagac gttgaaccct tgagggacct gggcaggggc ggtctgtggc tcctgctgga
     3481 acaactcaga gttagattgg cccccttgta actgcccgtc actaatactg tgcgtcaatg
     3541 tacgagagcc attcatacgg ctgccctcac ggccgtgatg ataaatgttt tggctttgta
     3601 agtcaaggtt aaggctcgtc atgtggttac gcaagcccga gatgccggac cgggaagcgg
     3661 ccgcacttcc cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga
     3721 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     3781 tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     3841 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     3901 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     3961 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     4021 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     4081 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     4141 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     4201 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     4261 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     4321 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     4381 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta
     4441 ccgtcgactg cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta
     4501 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     4561 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     4621 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     4681 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     4741 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     4801 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     4861 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     4921 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     4981 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5041 attactatta ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag
     5101 gtgggtctga aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac
     5161 agcaaaaact attcttaaac ctaccaagcc tcctactatc attatgaata attttatata
     5221 ccacagccaa tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa
     5281 taattcttgt tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa
     5341 gcttgtgtaa ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg
     5401 ttccagagat ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt
     5461 tccagagcaa ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag
     5521 gattcttgcc tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc
     5581 ctcaatagcc ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc
     5641 ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc
     5701 aaggaacaaa gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc
     5761 cttggtgggt gctactccta atggttcaat ttttactact ttatatttat ataattcact
     5821 tctccaattg tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta
     5881 atcctcatcc tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct
     5941 aatgatcttt gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac
     6001 tgccccttca cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc
     6061 cctgtaataa acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg
     6121 tatgtctgtt gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc
     6181 ttttctttta aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc
     6241 ctgatctctt acctgtccta taattttctt taattcttta ttcatagatt ctactactcc
     6301 ttgactttgg ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt
     6361 tttcttactt ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat
     6421 ctctatcctt tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc
     6481 ttctgatcct gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga
     6541 tgtttctaac aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat
     6601 atgttttaat ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga
     6661 tctaattctc ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc
     6721 gctagtcaaa atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc
     6781 ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc
     6841 cctgttcggg cgccactgct agagattttc cacactgact aaaagggtct gagggatctc
     6901 tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt
     6961 tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg
     7021 tctaaccaga gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc
     7081 tcgccactcc ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg
     7141 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     7201 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     7261 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     7321 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     7381 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     7441 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcgaccc
     7501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     7561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     7621 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7681 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7741 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7801 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7861 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7921 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7981 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     8041 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     8101 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     8161 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     8221 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     8281 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     8341 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     8401 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     8461 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     8521 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     8581 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     8641 agaaaagatc aaaggatctt c
//
