LOCUS       sequence-488242-        7488 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_012, derived from human protein PCDG1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7488)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7488)
  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_012
FEATURES             Location/Qualifiers
     source          1..7488
                     /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(2519..3229,3230..3232,3233..3235))
                     /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(2561..2582)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2910..2929
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3169..3190
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3232..3241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3317..3520)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3346..3366)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3521..3824
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4081..4125)
                     /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    4310..4543
                     /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    4903..5020
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5512..5637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5684..5941
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5943..6322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6353..6457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6458..6526,6527..7318)
                     /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(6676..6695)
                     /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 ccggttcact ttttttcctt tttacccgac ttctttttat
     2101 tgccgtttcc cccggctggg gctttaccgt cacgtttgcc agccgcgttt gttaatgtgg
     2161 cattacttcc cgggatgtaa acattttgac ggtagtcggg aacgtgttgc agcgtgaatt
     2221 gcggtccgta gcgagccgaa agccccattg tcccggctcc gccgccaagg gtacttgacc
     2281 catctgcagc ttcggatgca ctagcaagaa tcattgcttg caacatctcc gtgtcgaact
     2341 gattattagg ccatgttcca gtgtcatctc cgttttggct acctgaagtg ccagggcgtt
     2401 gggcctgtga gaagcgccaa tccgtattag gtggcgcttg ctgtgagaac ggttcctttt
     2461 tatcctccag cagtgaccgg gaagcggccg cacttccccc ggagcctccg cttccgccct
     2521 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     2581 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt
     2641 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     2701 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     2761 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     2821 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     2881 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     2941 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     3001 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     3061 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     3121 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     3181 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     3241 cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag
     3301 cccggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     3361 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     3421 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     3481 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     3541 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     3601 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     3661 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     3721 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     3781 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     3841 ggctatgaac taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct
     3901 tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     3961 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     4021 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     4081 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     4141 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     4201 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     4261 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     4321 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     4381 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     4441 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     4501 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     4561 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     4621 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg
     4681 tgttttacta aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc
     4741 acctgccatc tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt
     4801 tatgtcacta ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg
     4861 tcctttccaa agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
     4921 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     4981 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     5041 ttgtactgct gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa
     5101 ttctttattc atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc
     5161 tgaagatcct gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt
     5221 gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc
     5281 tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg
     5341 tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg
     5401 ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg
     5461 ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc
     5521 gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg
     5581 ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc
     5641 ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac
     5701 actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac
     5761 actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag
     5821 ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc
     5881 agctgcttat atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct
     5941 cccatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     6001 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     6061 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     6121 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     6181 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     6241 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     6301 taactagtca ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac
     6361 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6421 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6481 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6541 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6601 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6661 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6721 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6781 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6841 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6901 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6961 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7021 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7081 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7141 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7201 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7261 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7321 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7381 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7441 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
