LOCUS       sequence-488220-        7536 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_203, derived from human fusion protein TTC6_MIPOL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7536)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7536)
  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_203
FEATURES             Location/Qualifiers
     source          1..7536
                     /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(1069..1779,1780..1782,1783..1785))
                     /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(1111..1132)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1460..1479
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1719..1740
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1782..1791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1867..2070)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1896..1916)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2071..2374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2631..2675)
                     /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    2860..3093
                     /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    3453..3570
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4062..4187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4234..4491
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4493..4872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4903..5007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5008..5076,5077..5868)
                     /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(5226..5245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6039..6627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6528..6547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6781..6798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6866..7000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6890..6909)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6944..6963
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7006..7201)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7020..7155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7260..7281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7296..7301,7302..7319,7320..7326)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7334..7350
                     /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     7339..7361
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7358..7374
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7358..7374
                     /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 atccttcaga ttctcttctt ggctaaggga
      601 tttatgtaag tttttatgca gctttttgtc acaaagcggt tttgatgact tgcgcggacg
      661 caaactcttg cgcttgcgca tgcgcttgat ctccttcccc tcaatctgca gaaactcagg
      721 tgatttcgcc gtcagcgata cccatttcgc ctccttggag tcactagttg gctgtccgtc
      781 ctctaagggc tttgaaagtt cctctgctgg ttggacctgg gagatttcct gtggcttgaa
      841 cacgatagac tcggcttcac tcggtcccca ctcgatgtcc tcctgttcga tctgcaggat
      901 ggaggacatt gtctctgaca attctttatg aacactcatt tgtgactcct tggcccccat
      961 ttgaaacttg tattcggctt ggatttctgg cgtctccgct tgcatagggg acgtttttaa
     1021 atacatccgg gaagcggccg cacttccccc ggagcctccg cttccgccct tgtacagctc
     1081 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     1141 cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag
     1201 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     1261 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     1321 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     1381 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     1441 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     1501 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     1561 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     1621 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     1681 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     1741 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtgg
     1801 atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag cccggttcac
     1861 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     1921 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     1981 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     2041 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     2101 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     2161 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     2221 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     2281 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     2341 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     2401 taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct tgagctcggt
     2461 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     2521 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     2581 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     2641 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     2701 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     2761 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     2821 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     2881 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     2941 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     3001 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     3061 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     3121 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     3181 tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg tgttttacta
     3241 aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc acctgccatc
     3301 tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt tatgtcacta
     3361 ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg tcctttccaa
     3421 agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt aatttgtttt
     3481 tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc ccctgcactg
     3541 taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat ttgtactgct
     3601 gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa ttctttattc
     3661 atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc tgaagatcct
     3721 gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt gtctaaagct
     3781 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     3841 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     3901 tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc
     3961 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     4021 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     4081 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     4141 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     4201 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     4261 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     4321 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     4381 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
     4441 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct cccatggtaa
     4501 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4561 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4621 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4681 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4741 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4801 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     4861 ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4921 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4981 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5041 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5101 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5161 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5221 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5281 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5341 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5401 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5461 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5521 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5581 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5641 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5701 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5761 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5821 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5881 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5941 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6781 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6841 cgttggccga ttcattaatg cagcggatcc agacatgata agatacattg atgagtttgg
     6901 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6961 tgctttattt gtaaccatta taagctgcaa taaacaagtt agctttttgc aaaagcctag
     7021 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     7081 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     7141 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     7201 cggatccgct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7261 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7321 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7381 tacgccaagc ttgggctgca ggtcgaggga tctccataag agaagaggga cagctatgac
     7441 tgggagtagt caggagagga ggaaaaatct ggctagtaaa acatgtaagg aaaattttag
     7501 ggatgttaaa gaaaaaaata acacaaaaca aaatat
//
