LOCUS       sequence-488196-        7299 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_176, derived from human fusion protein SEMA6C_DLG2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7299)
  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_176
FEATURES             Location/Qualifiers
     source          1..7299
                     /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(832..1542,1543..1545,1546..1548))
                     /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(874..895)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1223..1242
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1482..1503
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1545..1554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1630..1833)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1659..1679)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1834..2137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2394..2438)
                     /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    2623..2856
                     /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    3216..3333
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3825..3950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3997..4254
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4256..4635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4666..4770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4771..4839,4840..5631)
                     /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(4989..5008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5802..6390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6291..6310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6544..6561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6629..6763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6653..6672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6707..6726
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6769..6964)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6783..6918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7023..7044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7059..7064,7065..7082,7083..7089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7097..7113
                     /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     7102..7124
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7121..7137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7121..7137
                     /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 cgctggctta tcgcaaagtt tatttacagt
      601 cgaatagact ggttctggtg ggcgggtata gtcgtcgtca accagcatgt gtttaggaat
      661 tgggctataa cgccccggag aaataggggg caacgaagtc ttgtactcaa gcgttccgtt
      721 gttaccacta agcaggtgat tctccatggg tggagaatat gaatgggtaa tgtcgggtgg
      781 cccgtagggc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc ccttgtacag
      841 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
      901 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg
      961 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     1021 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     1081 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     1141 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     1201 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     1261 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     1321 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     1381 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     1441 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     1501 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
     1561 tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac gagcccggtt
     1621 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     1681 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     1741 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     1801 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     1861 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     1921 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     1981 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2041 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     2101 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     2161 aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa gcttgagctc
     2221 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     2281 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     2341 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     2401 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     2461 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     2521 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     2581 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     2641 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     2701 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     2761 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     2821 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     2881 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     2941 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta tggtgtttta
     3001 ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc atcacctgcc
     3061 atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac ttttatgtca
     3121 ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc tggtcctttc
     3181 caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
     3241 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
     3301 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtact
     3361 gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt taattcttta
     3421 ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag gtctgaagat
     3481 cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat cttgtctaaa
     3541 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
     3601 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
     3661 atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa tcgttctagc
     3721 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
     3781 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
     3841 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
     3901 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
     3961 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
     4021 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     4081 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     4141 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
     4201 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctcccatgg
     4261 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4321 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4381 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4441 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4501 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4561 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4621 tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4981 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5281 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5941 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6481 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6541 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6601 gcgcgttggc cgattcatta atgcagcgga tccagacatg ataagataca ttgatgagtt
     6661 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     6721 tattgcttta tttgtaacca ttataagctg caataaacaa gttagctttt tgcaaaagcc
     6781 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     6841 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     6901 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     6961 atgcggatcc gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7021 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7081 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7141 gattacgcca agcttgggct gcaggtcgag ggatctccat aagagaagag ggacagctat
     7201 gactgggagt agtcaggaga ggaggaaaaa tctggctagt aaaacatgta aggaaaattt
     7261 tagggatgtt aaagaaaaaa ataacacaaa acaaaatat
//
