LOCUS       sequence-487925-        7293 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_006, derived from human fusion protein AFF1_RAD51B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7293)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7293)
  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_006
FEATURES             Location/Qualifiers
     source          1..7293
                     /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(826..1536,1537..1539,1540..1542))
                     /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(868..889)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1217..1236
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1476..1497
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1539..1548
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1624..1827)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1653..1673)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1828..2131
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2388..2432)
                     /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    2617..2850
                     /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    3210..3327
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3819..3944
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3991..4248
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4250..4629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4660..4764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4765..4833,4834..5625)
                     /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(4983..5002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5796..6384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6285..6304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6538..6555
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6623..6757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6647..6666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6701..6720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6763..6958)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6777..6912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7017..7038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7053..7058,7059..7076,7077..7083)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7091..7107
                     /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     7096..7118
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7115..7131
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7115..7131
                     /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 caccgggtgc tcgtccaagg tttctgtccg
      601 tatacccgtg gcccagttag taggagggag tgcagaactc aaaactgtag aatgcatctg
      661 cgggggacac atgcttactg agggcaacag gcggggattc ctggtctttc tcaaaacaac
      721 gtcagggata aggaatgttg gactgggatg tgaatcggcg gctgaccaac attcactccg
      781 cggccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
      841 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
      901 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
      961 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1021 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     1081 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1141 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1201 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1261 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1321 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     1381 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     1441 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     1501 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     1561 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     1621 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     1681 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     1741 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     1801 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     1861 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     1921 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     1981 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     2041 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     2101 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     2161 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     2221 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     2281 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     2341 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     2401 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     2461 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     2521 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     2581 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     2641 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     2701 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     2761 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     2821 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     2881 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     2941 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     3001 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     3061 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     3121 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     3181 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     3241 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     3301 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     3361 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     3421 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     3481 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     3541 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     3601 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     3661 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     3721 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     3781 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     3841 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     3901 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     3961 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     4021 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     4081 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     4141 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     4201 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     4261 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4321 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4381 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4441 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4501 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4561 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     4621 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4681 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4741 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4801 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4861 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4921 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4981 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5041 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5101 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5161 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5221 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5281 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5341 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5401 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5461 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5521 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5581 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5641 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5701 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5941 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6481 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6541 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6601 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     6661 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6721 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     6781 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     6841 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     6901 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     6961 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7021 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7081 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7141 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     7201 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     7261 tgttaaagaa aaaaataaca caaaacaaaa tat
//
