LOCUS       sequence-490659-        8718 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_013, derived from human fusion protein ATF7IP_JAK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8718)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8718)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_013
FEATURES             Location/Qualifiers
     source          1..8718
                     /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(2251..2961,2962..2964,2965..2967))
                     /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(2293..2314)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2642..2661
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2901..2922
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2964..2973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3049..3252)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3078..3098)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3253..3556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3813..3857)
                     /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    4042..4275
                     /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    4635..4752
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5244..5369
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5416..5673
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5675..6054
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6085..6189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6190..6258,6259..7050)
                     /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(6408..6427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7221..7809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7710..7729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7963..7980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8048..8182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8072..8091)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8126..8145
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8188..8383)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8202..8337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8442..8463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8478..8483,8484..8501,8502..8508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8516..8532
                     /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     8521..8543
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8540..8556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8540..8556
                     /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 ctttacttga aattctgcct catattcttc
      601 ttccatatat cgacgacgtt tagactgtac attgtccatg tcttctgatt tagaacgttt
      661 tcgtctagaa aattcatttt tctcagaagg tctttcatca tcttcatgta ttacttgctc
      721 ttcttcctca ggtctgttgt ccttttcatt actttctact tctgctggag agcacgtttc
      781 attttgcgat ataacttcag actcatcttt ttcacccgaa atatcctctt catcagagag
      841 gaccagaaag gcttcttttg gcaaactctc actactttct tgtttagatg gtgaaccaaa
      901 agaactttcc attttctctt ccaaatctgg atttgtctca tcgattggaa ggagagatgt
      961 tttagaaaag caagtaagtt catcctcaga aggtgcaagc ttttccaaaa taggaatgat
     1021 ttcatctgtc tccatagagt ctgtattttc taagatattc tcacttttgt tatcttcaat
     1081 aacattttca ttaatcgttt ctacaagatc tgcagaagtt tcatcttcat tcttttcacc
     1141 ttggtcaatt tccatactgc tagatatagc atcttctcca agagcaagct ctgtgataat
     1201 atcttcctct accttctttt cattttcagg aggtcgatct gaacaaatag ttgtatcatc
     1261 tgtttctaga gtttcttcat tagcatcaat attattatca gctttatttt tctcatccaa
     1321 tgagtcttta ctctgaattt gctctaattt ttcatcagct ccattttttt caagaaaatc
     1381 atcatcttta ttgctacttg gttctatatt gtcaatttca ggaactggtt cacaaactgg
     1441 tacagacttt ggttcaaagg tacgatcaaa ggttgattca gaagtcagct catcagaggc
     1501 cagttcacca gtggccagat catcagaggc cagttcacta gaggatagat caccagaggc
     1561 cagatcatca gaggctggat cagtggaagc tggtgctcca gaagccagat caacagaagt
     1621 tatttcacta gaggctatat catcagcggc acaatcacca gaaatgggtt caacagggac
     1681 cggttcacct gggatgggat cactagaggt gggatcacca gaggagagat catcagcagt
     1741 ggcatcacca gaggtggcat caccagggga cacatcacca gaaggggcat caccagaggt
     1801 tgcatcacca gaggcagcat cactagagga gggctcgcta gaggtgggat caccagaggt
     1861 ggaatcacca gaggccagta ctccggaggc tggatctccg gcatccagat caccaggggc
     1921 tggatcacca gaaactggtt cagcaggcag tttagagact ggttctggag ttatattatg
     1981 gggagacaaa ggttcacaat ttaaatcatc atcctgcttg ttttttacat taacacttag
     2041 gatacagggt gtttccttcc attctgcttt acttccttca ggatcaaaac aaatctcttc
     2101 aatgccattc acctcttcct tgtctttaat gtaatccatg ttttccaaag gtgaggttgg
     2161 gtccaaatct ccattttcat ggttgccatt taacagcttg acatcagtcc gggaagcggc
     2221 cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat
     2281 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     2341 gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     2401 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
     2461 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
     2521 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     2581 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     2641 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     2701 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     2761 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     2821 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     2881 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     2941 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg cccgcggtac
     3001 cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag ctctgcttat
     3061 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     3121 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     3181 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     3241 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     3301 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     3361 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     3421 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     3481 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     3541 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     3601 ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg
     3661 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
     3721 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
     3781 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
     3841 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
     3901 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
     3961 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
     4021 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
     4081 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
     4141 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
     4201 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
     4261 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
     4321 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
     4381 ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc catgttctaa
     4441 tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc ataatcccta
     4501 atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg tattactact
     4561 gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt tctgctgtcc
     4621 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
     4681 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
     4741 tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag atgttcagcc
     4801 tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc tactactcct
     4861 tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg ctgtgctttt
     4921 ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     4981 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     5041 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     5101 gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     5161 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat
     5221 ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg
     5281 ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct
     5341 tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc
     5401 ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg agggatctct
     5461 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     5521 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     5581 ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct
     5641 cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg actaatacgt
     5701 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     5761 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     5821 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     5881 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     5941 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     6001 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcgaccca
     6061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6901 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7921 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7981 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8041 tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     8101 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     8161 tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     8221 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     8281 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     8341 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg ctggcacgac
     8401 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8461 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8521 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgggctg
     8581 caggtcgagg gatctccata agagaagagg gacagctatg actgggagta gtcaggagag
     8641 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     8701 taacacaaaa caaaatat
//
