LOCUS       sequence-488262-        8739 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_033, derived from human protein IRS4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8739)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8739)
  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_033
FEATURES             Location/Qualifiers
     source          1..8739
                     /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"
     RBS             2382..2390
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(join(3770..4480,4481..4483,4484..4486))
                     /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(3812..3833)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4161..4180
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4420..4441
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4483..4492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4568..4771)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4597..4617)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4772..5075
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5332..5376)
                     /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    5561..5794
                     /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    6154..6271
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6763..6888
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6935..7192
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7194..7573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7604..7708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7709..7777,7778..8569)
                     /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(7927..7946)
                     /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 ccggttcacg cgatgatccc ccaactatca ggcaaccctt
     2101 gtgtactagg tgctggggta tttgactcac gctttgtgaa atccatgttt acatagtcac
     2161 ttgaggaatc tgcttcgcgc tgctcatgtg taggcttttg cggtttaggc ttaattttgt
     2221 aacctttcgt aatgaagcta aggcgatttg ggcgctttgc cttatttttg ggcggctcgt
     2281 ggtctgatgg tttcgaaggg gaaccgccgt cgcccggttt actaaacgag ccttccccgc
     2341 tcggtttgct tgcagcgtct tttgggtccc aattgtacga aacctcctta tccaaaccac
     2401 ggcccagaaa cttacccggc aacataggaa cgtactcgga attatcgttt tggcccaagg
     2461 gggagctgcg aaatgggttc ggaagtgaaa agtagctaga ccaggactta ctggaagatc
     2521 ctccctgtgg gttacgtggg tttttaggga tagcgcctgc acccggcgcc atgaacatat
     2581 agtccgattc ggagtcatta tctttcgagt catcctcttt gttggtatcc ggcgctttag
     2641 gtgggctggg tgcaggaggg gggctcacgc gaggaaacat catcatgtaa ccacgcgaat
     2701 cctcgaaagg cgaacgcgag tgacgttttt ttgacgcgga aacgttttgg ggggccatgg
     2761 gcatataatc agagctgctc acaaggggcg tggcaacacc cggacgcatg ggtacatagg
     2821 gatcgtcttc atcttcgtca aaagcacgag cgcggtgcgg tccgcgtgcg gcaccttctg
     2881 gaatctcggc gtctttaacc tctttggcct ccttacattc cttagtggct ccacgatcca
     2941 cacaaaaata taagcggaaa cggccaccgc tttttccctt gcctccagtg cccccagccg
     3001 gtgggggagg cggcggcgga ggcgggggtg gtggcatttg ttgttgcttt gattgcgtta
     3061 atttaccgaa gtagctacgc ttcttaagtg atttaccacg ctccccatca ccgtcggaac
     3121 ctttgcctga gccagaacct tttcctcctc cgctgttttt accccctccc gatccatggc
     3181 cgtctcccgg accttgacca ccacccgaac catgtccccc tcctgggcgc tgaccaccgc
     3241 cggaaccatg gcctccagct gtcccctgac cgtcacgtga acactgattt cctcctgagc
     3301 cttgaccgtt agatccttgg cccccgcggc tcccctgacc ttctccagag cattgattac
     3361 ctccggaaga gtggctggac gacccttgcc cgttggaacc ttgtcctcca cccgacccgc
     3421 ggccgtttcc agacccccaa ttattcattg gcatgtaatc ccctcccgac ccctcctgat
     3481 cttctttacc ttggggattg ccctcttcac caaaatttcc cgaacccgag ccagacactt
     3541 ccgacgataa acgtgcgcca ttattaggtg cctcagccgg atgacgtgga cgtgcaggag
     3601 acggagcaag acgacgaaag aagctagccg gtacggaaac cgcacgacgg gaacgacgac
     3661 cgcgaggaag gtgcagacgg ccgcggcggg aatgagcgac aggttcggag ggggtgacaa
     3721 agcggcgccg ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct
     3781 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     3841 gcttctcgtt ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca
     3901 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     3961 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     4021 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     4081 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     4141 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     4201 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     4261 ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca
     4321 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     4381 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     4441 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg
     4501 gatcccgggc ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca
     4561 ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     4621 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     4681 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     4741 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     4801 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     4861 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     4921 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     4981 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     5041 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     5101 ctaatgaccc cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg
     5161 tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct
     5221 attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat
     5281 tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc
     5341 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
     5401 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
     5461 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
     5521 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
     5581 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
     5641 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
     5701 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
     5761 tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg
     5821 caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt
     5881 atatttatat aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact
     5941 aaacttttcc atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat
     6001 ctgttttcca taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact
     6061 attatcttgt attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca
     6121 aagtggattt ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt
     6181 ttgtaattct ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact
     6241 gtacccccca atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc
     6301 tgtcttaaga tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt
     6361 catagattct actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc
     6421 tgctgcttgc tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc
     6481 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
     6541 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
     6601 ttgtctacag ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc
     6661 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
     6721 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
     6781 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
     6841 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt
     6901 ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa
     6961 aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga
     7021 agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc
     7081 tcccaggctc agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta
     7141 tatgcaggat ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta
     7201 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     7261 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     7321 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     7381 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     7441 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     7501 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     7561 aataatcaat gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7621 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7681 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7741 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7801 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7861 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7921 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7981 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     8041 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     8101 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     8161 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     8221 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     8281 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     8341 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     8401 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8461 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     8521 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     8581 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8641 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8701 ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
