LOCUS       sequence-488173-        7356 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_152, derived from human fusion protein PAX7_FOXO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7356)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7356)
  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_152
FEATURES             Location/Qualifiers
     source          1..7356
                     /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(889..1599,1600..1602,1603..1605))
                     /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(931..952)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1280..1299
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1539..1560
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1602..1611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1687..1890)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1716..1736)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1891..2194
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2451..2495)
                     /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    2680..2913
                     /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    3273..3390
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3882..4007
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4054..4311
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4313..4692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4723..4827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4828..4896,4897..5688)
                     /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(5046..5065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5859..6447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6348..6367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6601..6618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6686..6820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6710..6729)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6764..6783
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6826..7021)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6840..6975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7080..7101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7116..7121,7122..7139,7140..7146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7154..7170
                     /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     7159..7181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7178..7194
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7178..7194
                     /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 cgtcagtttt gtgcgttgcg ctaattcttc
      601 gcgcgtgtaa atgtcaggat aatgcgtacg ttcgaacgcc ttttccagtt cctccagctg
      661 ctctgccgta aatgtagtac gggagcggcg ttgtttgcgc ttcagcggaa ggtctggttc
      721 ggactcgaca tcgctgccct cgtcaaggcg gtttccttta tcccccaaaa taccatcaat
      781 tgaatgcttc gcctttttct ccccgtcgtc ttcttttttg tcggcctcat cctcctcctc
      841 tttcttccgg gaagcggccg cacttccccc ggagcctccg cttccgccct tgtacagctc
      901 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
      961 cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag
     1021 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     1081 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     1141 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     1201 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     1261 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     1321 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     1381 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     1441 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     1501 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     1561 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtgg
     1621 atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag cccggttcac
     1681 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     1741 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     1801 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     1861 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     1921 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     1981 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     2041 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     2101 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     2161 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     2221 taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct tgagctcggt
     2281 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     2341 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     2401 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     2461 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     2521 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     2581 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     2641 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     2701 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     2761 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     2821 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     2881 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     2941 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     3001 tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg tgttttacta
     3061 aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc acctgccatc
     3121 tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt tatgtcacta
     3181 ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg tcctttccaa
     3241 agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt aatttgtttt
     3301 tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc ccctgcactg
     3361 taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat ttgtactgct
     3421 gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa ttctttattc
     3481 atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc tgaagatcct
     3541 gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt gtctaaagct
     3601 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     3661 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     3721 tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc
     3781 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     3841 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     3901 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     3961 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     4021 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     4081 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     4141 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     4201 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
     4261 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct cccatggtaa
     4321 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4381 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4441 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4501 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4561 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4621 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     4681 ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4801 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4921 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4981 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5041 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5101 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5221 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5281 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5401 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5701 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5761 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5821 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5881 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5941 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6001 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6061 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6121 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6181 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6241 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6301 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6361 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6421 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6481 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6541 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6601 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6661 cgttggccga ttcattaatg cagcggatcc agacatgata agatacattg atgagtttgg
     6721 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6781 tgctttattt gtaaccatta taagctgcaa taaacaagtt agctttttgc aaaagcctag
     6841 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6901 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6961 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     7021 cggatccgct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7081 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7141 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7201 tacgccaagc ttgggctgca ggtcgaggga tctccataag agaagaggga cagctatgac
     7261 tgggagtagt caggagagga ggaaaaatct ggctagtaaa acatgtaagg aaaattttag
     7321 ggatgttaaa gaaaaaaata acacaaaaca aaatat
//
