LOCUS       sequence-488240-        7794 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_010, derived from human protein FOXN3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7794)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7794)
  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_010
FEATURES             Location/Qualifiers
     source          1..7794
                     /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(2825..3535,3536..3538,3539..3541))
                     /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(2867..2888)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3216..3235
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3475..3496
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3538..3547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3623..3826)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3652..3672)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3827..4130
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4387..4431)
                     /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    4616..4849
                     /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    5209..5326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5818..5943
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5990..6247
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6249..6628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6659..6763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6764..6832,6833..7624)
                     /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(6982..7001)
                     /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 ccggttcaat tcttcgtcgt ttccttctgt tctttctggc
     2101 ccttcgccgt gcgattagtg atattgttta agcatgaacg aatcccagcc aaatgcagaa
     2161 ggctccccgc tgcttccttc atctcttcgt cgtctgattc aggaggcttt tcagtacgac
     2221 gtttcttcaa aggtaaggtg tcgctgggta ccttgcgtgc ttttgcaaag tgttgacgtt
     2281 ttttatgctg gctagcatat ccagaatccc ccaaactatc cttcggctct ttttgcgagt
     2341 gcttacggtc gtcctcctct gtgtcgctgg gtgactcatg cgagcgaaac gatccttcac
     2401 tcccctcgct tccttcttga ctacccttag tagcaaactc gtagtggtcg tcagcagagc
     2461 tgctcgaact actaatcgaa tcgctcgtgg gcgaggtaga acgggcatta ctcgatttgg
     2521 cagaggagta attatggtct tctttagggt cgccactgac cacaggcgat ccacaagagg
     2581 gttccgattc agtgcgcata cgacaggatg taatcccgtt gcgcatggct gcggtaacac
     2641 cgatcggcgt gataggcaaa ggacgaacac ctggaaacag tccacgggat aaaacacgtg
     2701 cgccgttctg gatgactccc ggcgggaacc ctgcctggat ctcaggcggg gtattcagca
     2761 gcatcatagc cgatgccgca tcccgggaag cggccgcact tcccccggag cctccgcttc
     2821 cgcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     2881 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg gtgctcaggt
     2941 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     3001 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc
     3061 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     3121 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     3181 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     3241 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     3301 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     3361 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     3421 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     3481 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     3541 tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga ctgcagaatt cggttcacta
     3601 aacgagcccg gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac
     3661 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     3721 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     3781 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     3841 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     3901 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     3961 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4021 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4081 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4141 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gcacgcgttc
     4201 gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
     4261 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
     4321 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
     4381 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
     4441 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
     4501 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
     4561 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
     4621 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     4681 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     4741 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     4801 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     4861 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     4921 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     4981 ttatggtgtt ttactaaact tttccatgtt ctaatcctca tcctgtctac ttgccacaca
     5041 atcatcacct gccatctgtt ttccataatc cctaatgatc tttgcttttc ttcttggcac
     5101 tacttttatg tcactattat cttgtattac tactgcccct tcacctttcc agaggagctt
     5161 tgctggtcct ttccaaagtg gatttctgct gtccctgtaa taaacccgaa aattttgaat
     5221 ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat
     5281 tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt ggatgaatac
     5341 tgccatttgt actgctgtct taagatgttc agcctgatct cttacctgtc ctataatttt
     5401 ctttaattct ttattcatag attctactac tccttgactt tggggattgt agggaatttc
     5461 caggtctgaa gatcctgctg cttgctgtgc ttttttctta cttttgtttt gctcttcctc
     5521 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag
     5581 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg
     5641 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag tattaactgc
     5701 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc
     5761 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac
     5821 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac
     5881 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga
     5941 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt
     6001 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     6061 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     6121 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca
     6181 aaaagcagct gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc
     6241 acgcctccca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6301 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6361 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     6421 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     6481 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     6541 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     6601 tattaataac tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg
     6661 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6721 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6781 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6841 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6901 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6961 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7021 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7081 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7141 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7201 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7261 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7321 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7381 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7441 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7501 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7561 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7621 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7681 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7741 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
