LOCUS       sequence-487983-        7386 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_074, derived from human fusion protein ITSN2_ADCY3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7386)
  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_074
FEATURES             Location/Qualifiers
     source          1..7386
                     /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(919..1629,1630..1632,1633..1635))
                     /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(961..982)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1310..1329
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1569..1590
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1632..1641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1717..1920)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1746..1766)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1921..2224
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2481..2525)
                     /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    2710..2943
                     /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    3303..3420
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3912..4037
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4084..4341
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4343..4722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4753..4857
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4858..4926,4927..5718)
                     /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(5076..5095)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5889..6477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6378..6397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6631..6648
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6716..6850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6740..6759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6794..6813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6856..7051)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6870..7005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7110..7131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7146..7151,7152..7169,7170..7176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7184..7200
                     /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     7189..7211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7208..7224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7208..7224
                     /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 ttcgcgttgc ttacgttccc attcctcctt
      601 ctccttttgt gccttacgct cagcctcacg ctgctgttgt tccatcaagg cttgacgacg
      661 tttttccagt tccatattcc cgcgctcata gttggccttg cgtttatctt cgaacgttac
      721 gggcagcttc ttttggggtt cctcctcctg cattttttgg taagagggaa gggtaccatt
      781 gatactatcg atttgtttac ccccacggaa gctgggagga accaattccg gcggcaatgt
      841 aagaggaagg ggctgcccag ctttagccat atcggtccgg gaagcggccg cacttccccc
      901 ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
      961 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     1021 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1081 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     1141 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     1201 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1261 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     1321 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     1381 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     1441 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     1501 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     1561 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     1621 gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag
     1681 aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat agacctccca
     1741 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     1801 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     1861 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     1921 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     1981 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2041 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2101 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2161 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2221 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     2281 actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac
     2341 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     2401 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     2461 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     2521 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     2581 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     2641 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     2701 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     2761 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     2821 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     2881 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     2941 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     3001 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     3061 ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc ctcatcctgt
     3121 ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat gatctttgct
     3181 tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc cccttcacct
     3241 ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct gtaataaacc
     3301 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
     3361 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
     3421 ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg atctcttacc
     3481 tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg actttgggga
     3541 ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt cttacttttg
     3601 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     3661 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     3721 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     3781 ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     3841 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     3901 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     3961 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     4021 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     4081 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     4141 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     4201 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     4261 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     4321 gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag atgtactgcc
     4381 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4441 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4501 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4561 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4621 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4681 ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt
     4741 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4801 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4861 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4921 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4981 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5041 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5101 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5161 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5221 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5281 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5341 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5401 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5461 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5521 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5581 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5641 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5701 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5761 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5821 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5881 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5941 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6001 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6061 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6121 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6181 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6241 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6301 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6361 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6421 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6481 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6541 tttcctgcgt tatcccctgc ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6601 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6661 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     6721 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     6781 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     6841 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     6901 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     6961 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     7021 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     7081 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7141 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7201 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     7261 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     7321 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     7381 aaatat
//
