LOCUS       sequence-488180-        7287 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_160, derived from human fusion protein RABGAP1L_BCAS3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7287)
  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_160
FEATURES             Location/Qualifiers
     source          1..7287
                     /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(820..1530,1531..1533,1534..1536))
                     /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(862..883)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1211..1230
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1470..1491
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1533..1542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1618..1821)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1647..1667)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1822..2125
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2382..2426)
                     /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    2611..2844
                     /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    3204..3321
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3813..3938
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3985..4242
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4244..4623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4654..4758
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4759..4827,4828..5619)
                     /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(4977..4996)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5790..6378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6279..6298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6532..6549
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6617..6751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6641..6660)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6695..6714
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6757..6952)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6771..6906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7011..7032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7047..7052,7053..7070,7071..7077)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7085..7101
                     /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     7090..7112
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7109..7125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7109..7125
                     /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 ggcatcgatc actgtactaa cgccgcggcg
      601 gtcgctgaca ggacgagaac ttccgggcat gcttacaggg tccgagcgga ctggctgaat
      661 gcgaagacta ttcaaatcaa ggtcgtcggt atcaaaatcc agcagcggtt gaggggtgtc
      721 tgaagggcca tgcgactgta atacggagga actggaacta atgactgtgg tttgcccccg
      781 ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct cgtccatgcc
      841 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
      901 ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
      961 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     1021 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     1081 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     1141 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     1201 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     1261 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     1321 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     1381 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     1441 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     1501 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg gatcccgggc
     1561 ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca ctaaacgagc
     1621 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     1681 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     1741 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     1801 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     1861 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     1921 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     1981 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     2041 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     2101 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     2161 cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg tcccctcggg
     2221 gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata
     2281 gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat
     2341 tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct
     2401 aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag
     2461 tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc
     2521 caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa
     2581 atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc
     2641 acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc
     2701 tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat
     2761 tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc
     2821 aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt
     2881 ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat
     2941 aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact aaacttttcc
     3001 atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat ctgttttcca
     3061 taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact attatcttgt
     3121 attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca aagtggattt
     3181 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
     3241 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
     3301 atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc tgtcttaaga
     3361 tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt catagattct
     3421 actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc tgctgcttgc
     3481 tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg
     3541 tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat
     3601 ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag
     3661 ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc cctgcttgcc
     3721 catactatat gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc
     3781 catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc
     3841 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
     3901 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
     3961 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
     4021 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
     4081 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
     4141 agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta tatgcaggat
     4201 ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta atagcgatga
     4261 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     4321 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     4381 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     4441 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     4501 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     4561 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     4621 gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4681 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4741 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4801 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4861 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4921 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4981 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5041 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5101 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5161 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5221 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5281 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5341 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5401 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5461 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5521 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5581 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5641 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5701 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6421 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6481 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6541 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6601 attcattaat gcagcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     6661 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     6721 tgtaaccatt ataagctgca ataaacaagt tagctttttg caaaagccta ggcctccaaa
     6781 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     6841 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     6901 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcggatccgc
     6961 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7021 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7081 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7141 cttgggctgc aggtcgaggg atctccataa gagaagaggg acagctatga ctgggagtag
     7201 tcaggagagg aggaaaaatc tggctagtaa aacatgtaag gaaaatttta gggatgttaa
     7261 agaaaaaaat aacacaaaac aaaatat
//
