LOCUS       sequence-487924-        8106 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_005, derived from human fusion protein AFF1_RAD51B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8106)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8106)
  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_005
FEATURES             Location/Qualifiers
     source          1..8106
                     /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(1639..2349,2350..2352,2353..2355))
                     /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(1681..1702)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2030..2049
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2289..2310
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2352..2361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2437..2640)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2466..2486)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2641..2944
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3201..3245)
                     /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    3430..3663
                     /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    4023..4140
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4632..4757
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4804..5061
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5063..5442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5473..5577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5578..5646,5647..6438)
                     /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(5796..5815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6609..7197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7098..7117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7351..7368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7436..7570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7460..7479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7514..7533
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7576..7771)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7590..7725
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7830..7851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7866..7871,7872..7889,7890..7896)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7904..7920
                     /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     7909..7931
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7928..7944
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7928..7944
                     /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 cagctttgcc tttgccggca ccttgagatc
      601 tgttttctcg aaggtctgtt gtctatagtc ttcggggagc ggcttcaact cagggctttc
      661 tgatggggcc tggtcctgac cgtccatcgg ccttacgtat gctgttggct tttgctgcat
      721 cgcgacagac tttgatggca gagatggggg tggaaatgtt tgggatggtg gctgggcagg
      781 ggcaacgagg gagtcctgtg gtgtctcttt atcatgtacc ttgaccgcca gatctttagg
      841 agactttgcc gggcaatatc ctttgctatt gtttgaggag ccgtgcacct ttgagcttcc
      901 ctgagtgcgg ggaagcgtct gttggttgga gtggatggga gagagtggtg gaacggggga
      961 cggcaagctg ataagaggag agagttcgcg ctctggcgca ctatccgtaa cggaagcgca
     1021 gtggtctccg tctgctcttc tatctccctt tttgtggtga ctggaaccaa atccttcctg
     1081 gccgagtcga tcttgggtca ggtgctgact atctgggggc ccgcaggatt ttgcgtggag
     1141 tgatggcatc ggttccgtgc ggggctgggc cattttggga ttatgagaaa tgttacctac
     1201 agacaatggg ccggatgcag gggtgtgaat gctctgatga tgtacactcg tgtgaaatga
     1261 gcttgacggg atgctgctgc ccttatcagg tatcaagggg tatttaggtt tcccgaggcg
     1321 attttcactg gcgtcaaggc ggtgggtgtg actcttagtg ctcaggaact cctttacttc
     1381 ctcataatta cccaacatgt tttgtattcg ggaactcaat tcatctccct tggctgtctt
     1441 ataaggctct ccgaagaggg gaattttttc agggaatgcc tccttctctt ggtgagcttc
     1501 ctgattgcgc cgttcctttt ctcgtattct gagcaagttc cgatcatcgt tataaagact
     1561 gttcccggat gagttcactc tctcagtaaa agccatccgg gaagcggccg cacttccccc
     1621 ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1681 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     1741 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1801 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     1861 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     1921 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1981 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     2041 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     2101 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     2161 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     2221 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     2281 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     2341 gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag
     2401 aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat agacctccca
     2461 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     2521 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2581 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2641 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2701 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2761 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2821 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2881 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2941 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3001 actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac
     3061 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     3121 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     3181 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     3241 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     3301 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     3361 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     3421 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     3481 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     3541 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     3601 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     3661 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     3721 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     3781 ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc ctcatcctgt
     3841 ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat gatctttgct
     3901 tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc cccttcacct
     3961 ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct gtaataaacc
     4021 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
     4081 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
     4141 ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg atctcttacc
     4201 tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg actttgggga
     4261 ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt cttacttttg
     4321 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     4381 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     4441 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     4501 ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     4561 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     4621 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     4681 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     4741 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     4801 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     4861 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     4921 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     4981 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     5041 gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag atgtactgcc
     5101 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     5161 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     5221 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     5281 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     5341 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     5401 ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt
     5461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5581 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5641 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5701 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5761 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5821 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5881 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5941 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6001 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6061 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6121 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6181 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6241 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6301 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6361 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6421 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7261 tttcctgcgt tatcccctgc ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7321 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7381 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     7441 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     7501 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     7561 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     7621 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     7681 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     7741 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     7801 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7861 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7921 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     7981 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     8041 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     8101 aaatat
//
