LOCUS       sequence-488223-        8097 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_208, derived from human fusion protein VAV2_COL27A1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8097)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8097)
  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_208
FEATURES             Location/Qualifiers
     source          1..8097
                     /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(1630..2340,2341..2343,2344..2346))
                     /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(1672..1693)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2021..2040
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2280..2301
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2343..2352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2428..2631)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2457..2477)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2632..2935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3192..3236)
                     /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    3421..3654
                     /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    4014..4131
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4623..4748
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4795..5052
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5054..5433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5464..5568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5569..5637,5638..6429)
                     /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(5787..5806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6600..7188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7089..7108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7342..7359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7427..7561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7451..7470)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7505..7524
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7567..7762)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7581..7716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7821..7842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7857..7862,7863..7880,7881..7887)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7895..7911
                     /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     7900..7922
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7919..7935
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7919..7935
                     /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 gctgtagctc tccggacgca gcgcaccgct
      601 ggtatccatc caggtctgaa atgcggcgcc gagatcgtct tgttgaagtt ggatcggacc
      661 acccggcgga cctggcggac ctggtctacc gcgaggacct ggcgggccgc gcgggccttg
      721 caaaccccag tcgccacgag atcctttgtc acccttcggc cccggcagac cactagggcc
      781 caaaatgccc aacgggccga cgatgccctc tttacccatc aggcccggcg gaccacgcgg
      841 cccctggatg cctttgaaac cgatatcgcc cggaggtccc tttgggccga ataaacctgc
      901 catgcccggg aaacccatct cgccgctgtc gcccggctga cctttggagc caggctctcc
      961 ctggttaccc gggagaccgg tacgcccttc cgtacctctc ttgcccggag gacccaattg
     1021 acccggcaaa ccggactcac ccttgaagcc cggcggaccc tgcgcgcccg gcaggcctgc
     1081 gacacccgga ttaccgcgct tgcccgctgg tcccatcggg cccgggtcgc cgtcatcacc
     1141 ttgctcacct tgttgaccag cttggccgtc tctgcctggc aggccgtcgg ggcccgcgat
     1201 gccttccaga ccctggcggc ccacaacgcc acgcggaccc ggcagaccct gcagaccttg
     1261 cacgccacgg cgccccggag cacccgcctt tccctgctta cctttcgggc cttcagcgcc
     1321 tttggaaccc ttcggacccg gccagccacg cgggcccggg tgaccgggtt gaccttgttg
     1381 acccggttta ccacgcaggc cctgcacacc ttcttggccc ggataacccg gatcacccgg
     1441 ttcaccacga tcgccacgat ctcctaccgg accgcggtcg cccggcggac ctggtgggcc
     1501 ttccgccttc ccgtcttcgc cttgctcgcc tttctcgccc ttcgggcccg gcggacccaa
     1561 cggacccacg atgcctttgt gacctttccg ggaagcggcc gcacttcccc cggagcctcc
     1621 gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1681 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     1741 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     1801 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     1861 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     1921 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     1981 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     2041 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     2101 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     2161 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     2221 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     2281 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     2341 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggtt
     2401 cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2461 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2521 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2581 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2641 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2701 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2761 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2821 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2881 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     2941 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagcacg
     3001 cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     3061 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     3121 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     3181 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     3241 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     3301 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     3361 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     3421 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     3481 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     3541 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     3601 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     3661 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     3721 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     3781 cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg tctacttgcc
     3841 acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc ttttcttctt
     3901 ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc tttccagagg
     3961 agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac ccgaaaattt
     4021 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     4081 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     4141 aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac ctgtcctata
     4201 attttcttta attctttatt catagattct actactcctt gactttgggg attgtaggga
     4261 atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt gttttgctct
     4321 tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa
     4381 tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat
     4441 ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag gccagtatta
     4501 actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt atattttttc
     4561 tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata
     4621 ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta
     4681 ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt
     4741 cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag
     4801 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     4861 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     4921 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     4981 aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc
     5041 aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     5101 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     5161 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     5221 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     5281 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     5341 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5401 attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt tcggggaaat
     5461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6721 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7261 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7321 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7381 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcggatc cagacatgat
     7441 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     7501 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     7561 tagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7621 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7681 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7741 ggttgctgac taattgagat gcggatccgc tggcacgaca ggtttcccga ctggaaagcg
     7801 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7861 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7921 ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa
     7981 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     8041 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatat
//
