LOCUS       sequence-488197-        7758 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_177, derived from human fusion protein SLC16A14_SP110.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7758)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7758)
  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_177
FEATURES             Location/Qualifiers
     source          1..7758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(741..760)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    806..861
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(860..879)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1222..1238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1222..1238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1235..1257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1246..1262
                     /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)."
     promoter        complement(join(1270..1276,1277..1294,1295..1300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1315..1336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1395..1590
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1441..1576
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1596..1730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1633..1652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1687..1706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1798..1815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1969..2557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2049..2068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2728..3519,3520..3588))
                     /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     3351..3370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3589..3693)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3724..4103
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             4105..4362
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4409..4534
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5026..5143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5503..5736
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5921..5965
                     /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"
     enhancer        6222..6525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6526..6729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6680..6700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6805..6814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6811..6813,6814..6816,6817..7527)
                     /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(6856..6877)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7117..7136)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7464..7485
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaaaa gtttaccggg cgggactgca tcttcccgtc atggtatcca gaaaaagttg
       61 aagcgcgtag accaggtccc ccaaaagaag gatgacagca cttgcaattc caccgtggag
      121 actcgtgcgc agaaagctcg cactgagtgc gcacgcaaga gtcgttcgga agaaattatc
      181 gatggtacat ccgaaatgaa tgagggcaag cgcagccaga agaccccgtc cacaccacgt
      241 cgcgtgaccc aaggggctgc ctcaccgggg catggaatcc aggaaaaact tcaggtcgtt
      301 gacaaggtca cacagcgtaa agacgactcg acgtggaata gtgaggtgat gatgcgtgtc
      361 caaaaagccc gcaccaagtg cgctcgcaag tcacgtttaa aagagaagaa gaaagaaaaa
      421 gacatctgtt cctcatccaa gcgccgtttt cagaagaaca tccaccgtcg tggcaagcct
      481 aagtcagatt gaaccggttc tagataagat tacggtacct ttaagaccaa tgacttacaa
      541 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
      601 ctcccaaaga agacaagatc tgctttttgc ctgtactggg tctctctggt tagaccagat
      661 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      721 cagctgctcg agctagcaga tctttttccc tctgccaaaa attatgggga catcatgaag
      781 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
      841 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
      901 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
      961 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     1021 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     1081 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     1141 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     1201 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1381 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     1441 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     1501 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     1561 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     1621 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1681 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     1741 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1801 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1861 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1921 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1981 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2041 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2101 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2161 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2221 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2281 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2341 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2401 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2461 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2521 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2581 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2641 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2701 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2761 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2821 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2881 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2941 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3001 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3061 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3121 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3181 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3241 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3301 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3361 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3421 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3481 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3541 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3601 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3661 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     3721 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3781 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3841 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3901 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3961 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4021 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4081 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
     4141 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     4201 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     4261 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     4321 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     4381 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     4441 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     4501 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     4561 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     4621 agtatgggca agcagggagc tagaacgatt cgcagttaat actggcctgt tagaaacatc
     4681 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     4741 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     4801 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     4861 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     4921 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     4981 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     5041 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     5101 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     5161 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     5221 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     5281 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     5341 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     5401 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     5461 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     5521 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     5581 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     5641 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     5701 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     5761 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     5821 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     5881 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     5941 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     6001 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     6061 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     6121 cccaaccccg aggggaccga gctcaagctt cgaacgcgtg ctagttatta atagtaatca
     6181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     6241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     6301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     6541 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     6601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6661 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6721 agcagagctc gtttagtgaa ccgggctcgt ttagtgaacc gaattctgca gtcgacggta
     6781 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     6841 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     6901 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     6961 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     7021 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     7081 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     7141 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     7201 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     7261 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     7321 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     7381 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     7441 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     7501 actctcggca tggacgagct gtacaagggc ggaagcggag gctccggggg aagtgcggcc
     7561 gcttcccggc cacaaatccg cgataaagaa gaccctcaag agatgcctca ttccccctta
     7621 ggttctatgc ctgagatccg cgacaatagt ccggaaccca atgacccgga agagccccag
     7681 gaggtaagct cgaccccaag cgataagaag gggaagaagc gtaagcgctg tatctggagc
     7741 acgcccaagc gccgccat
//
