LOCUS       sequence-488178-        7701 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_158, derived from human fusion protein PTK2_PSMA7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7701)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7701)
  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_158
FEATURES             Location/Qualifiers
     source          1..7701
                     /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(1234..1944,1945..1947,1948..1950))
                     /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(1276..1297)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1625..1644
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1884..1905
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1947..1956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2032..2235)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2061..2081)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2236..2539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2796..2840)
                     /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    3025..3258
                     /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    3618..3735
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4227..4352
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4399..4656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4658..5037
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5068..5172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5173..5241,5242..6033)
                     /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(5391..5410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6204..6792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6693..6712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6946..6963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7031..7165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7055..7074)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7109..7128
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7171..7366)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7185..7320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7425..7446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7461..7466,7467..7484,7485..7491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7499..7515
                     /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     7504..7526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7523..7539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7523..7539
                     /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 agaatccgca ggactggata aactcgctaa
      601 ggaacccaga tgaccggggg ctcccggacg tggcggcttt ttcggaggtg ccgccggatc
      661 gggtttcccc accggttggt aaatatgctg attcccaatg ggtccctgca aggacccatc
      721 ctcgcggtcg atactcccac ggctaagacg cacatctggc ttcagaaaac gctcttcttt
      781 ttctaaccaa cgctgatctt cctccatttc ctgttgctga cggatcagac gttcctccat
      841 aagatgagtc ggcagtacct gaccaattcc gcgaaggtcc agcacagtcg agtcttcaac
      901 gtttggctgc cacatcgcga tctcctgtgg gcggtgattc catgaatctg tttgatctaa
      961 caaactcgcc tgtccagggt aaattgaccc tgccatcgcc gtaataccgt gactgccagg
     1021 atacccggag acttggtaat ggttagtttg aaccatgtgc tggggggagg ggtaaaagcc
     1081 ctcgctactg cgcggggatg ggtagcccgg acgtgatggc tttggagggg cttcatcaga
     1141 tccacccgaa tcccaagaga cagtggcttg gcgacggctc tccatacgca tccgggaagc
     1201 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     1261 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1321 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1381 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1441 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1501 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1561 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1621 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     1681 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     1741 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1801 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     1861 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1921 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     1981 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     2041 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     2101 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2161 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2221 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2281 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2341 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2401 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2461 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2521 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2581 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     2641 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     2701 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     2761 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     2821 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     2881 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     2941 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     3001 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     3061 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     3121 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     3181 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     3241 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     3301 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     3361 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     3421 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     3481 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     3541 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     3601 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     3661 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     3721 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     3781 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     3841 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     3901 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     3961 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     4021 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     4081 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     4141 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     4201 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     4261 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     4321 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     4381 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     4441 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     4501 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     4561 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     4621 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     4681 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4741 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4801 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4861 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4921 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4981 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     5041 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5101 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5161 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5221 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5281 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5341 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5401 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5461 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5521 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5581 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5641 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5701 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5761 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5821 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5881 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5941 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6001 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6061 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6121 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6841 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6901 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6961 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7021 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7081 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7141 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     7201 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7261 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7321 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     7381 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7441 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7501 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     7561 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     7621 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     7681 aaataacaca aaacaaaata t
//
