LOCUS       sequence-488260-        8394 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_031, derived from human protein MD12L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8394)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8394)
  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_Vidr_031
FEATURES             Location/Qualifiers
     source          1..8394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3425..4135,4136..4138,4139..4141))
                     /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(3467..3488)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3816..3835
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4075..4096
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4138..4147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4223..4426)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4252..4272)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4427..4730
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4987..5031)
                     /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    5216..5449
                     /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    5809..5926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6418..6543
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6590..6847
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6849..7228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7259..7363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7364..7432,7433..8224)
                     /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(7582..7601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct gctagctcga gcagctgaag ctttattgag gcttaagcag
     1861 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
     1921 agtacaggca aaaagcagat cttgtcttct ttgggagtga attagccctt ccagtccccc
     1981 cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg
     2041 taccgtaatc ttatctagaa ccggttcaga aatgagatgg atgcccgtat ggagttaccc
     2101 cttgctgagg ctggttggag cttaactgtt tctgcagttg acgaaccagc gcagctgtct
     2161 gctgttgttg ctgagtctgc tgcagcccct gacgtgggaa taacggttgc tgtggttgca
     2221 tagcttgcaa accaagtgtt tgactttggg attggcttga ttgctgcggt tgtgggggct
     2281 gttggggctg cggctgctgg ggctgctgca tttgaagcag gcgctgttgt tggcggacct
     2341 gtggttggcg ggggcgcatg gggtcttgag gtaacggaac ggaaggaagg ttagggtgtg
     2401 ccgaagttaa gacagcgtca atgcctccac cgacaagggg gctctgctga agggcctgat
     2461 gattcaggcg ttgactccca gtcagtggtt gaagatacgg tgatgcctgt tgttgcacat
     2521 agcctgaggg ctgctgttga atttggcgaa gacgttccat caatacggga tttgaatgcg
     2581 ccgcagggta tgcgcgagaa ttgtagcttg ggcttaagac cacagatccc gcttgctgtt
     2641 gggatgtctg ctgcagcggc atctgggttc cgtacatggt gtatccttgc ggcattgtct
     2701 gagcttgttg caagccggga tattggggca gttgaggcga tggacgagcc tgtgctgcaa
     2761 aaagagctgc ttgatcgccc ggctgccctt gttggaacgg gcgagtttgt gcttggcgaa
     2821 gtaagcgttg ttgttgttgc tgctgcaaca atttcatttg gatcaattgt tgctgcgagg
     2881 tgatagcgtg aaggctcggc ggctgcatca tggctcctga gcggcgctgt aacatgtttg
     2941 ataaagcctg cttcgtattt gtaggtacga acgagccggc aggatctaag cgtgagcccc
     3001 caggagtcag actctgattc tgcaaaaaaa aaccgggctg ttgtggttga ccaaccaggt
     3061 tataacccca taaggtggat tgtggatgat gcatcatctg agagctaata gggctataat
     3121 tggggggtac acgataaata ttagactgcg ggtattcatc aacgcgtgaa ctagacttag
     3181 ttttgcgctt gcgacctttg gtttttttct cctcatctgt ggtagttttc ccttgatcag
     3241 acaactccgc agatttgcgt tcaggttcct gggaaaccgg gctagtaggt tcctcctctt
     3301 cttcctctgg cggaagaggc aacggttgca agtaatagct acggggtttt ggcattggat
     3361 gcgtatggta cagaagcaaa tgccgggaag cggccgcact tcccccggag cctccgcttc
     3421 cgcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     3481 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg gtgctcaggt
     3541 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     3601 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc
     3661 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     3721 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     3781 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     3841 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     3901 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     3961 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     4021 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     4081 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     4141 tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga ctgcagaatt cggttcacta
     4201 aacgagcccg gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac
     4261 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     4321 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     4381 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     4441 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4501 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4561 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4621 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4681 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4741 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gcacgcgttc
     4801 gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
     4861 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
     4921 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
     4981 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
     5041 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
     5101 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
     5161 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
     5221 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     5281 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     5341 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     5401 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     5461 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     5521 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     5581 ttatggtgtt ttactaaact tttccatgtt ctaatcctca tcctgtctac ttgccacaca
     5641 atcatcacct gccatctgtt ttccataatc cctaatgatc tttgcttttc ttcttggcac
     5701 tacttttatg tcactattat cttgtattac tactgcccct tcacctttcc agaggagctt
     5761 tgctggtcct ttccaaagtg gatttctgct gtccctgtaa taaacccgaa aattttgaat
     5821 ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat
     5881 tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt ggatgaatac
     5941 tgccatttgt actgctgtct taagatgttc agcctgatct cttacctgtc ctataatttt
     6001 ctttaattct ttattcatag attctactac tccttgactt tggggattgt agggaatttc
     6061 caggtctgaa gatcctgctg cttgctgtgc ttttttctta cttttgtttt gctcttcctc
     6121 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag
     6181 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg
     6241 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag tattaactgc
     6301 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc
     6361 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac
     6421 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac
     6481 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga
     6541 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt
     6601 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     6661 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     6721 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca
     6781 aaaagcagct gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc
     6841 acgcctccca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6901 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6961 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     7021 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     7081 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     7141 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     7201 tattaataac tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg
     7261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
