LOCUS       sequence-488267-        8424 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged fusion protein PAX5_ZCCHC7 with an IDR 
            deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8424)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8424)
  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_PAX5_ZCCHC7-Delta-1-447
FEATURES             Location/Qualifiers
     source          1..8424
                     /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(1957..2667,2668..2670,2671..2673))
                     /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(1999..2020)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2348..2367
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2607..2628
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2670..2679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2755..2958)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2784..2804)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2959..3262
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3519..3563)
                     /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    3748..3981
                     /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    4341..4458
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4950..5075
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5122..5379
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5381..5760
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5791..5895
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5896..5964,5965..6756)
                     /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(6114..6133)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6927..7515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7416..7435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7669..7686
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7754..7888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7778..7797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7832..7851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7894..8089)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7908..8043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8148..8169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8184..8189,8190..8207,8208..8214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8222..8238
                     /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     8227..8249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8246..8262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8246..8262
                     /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 ctcgcgctct tggatttcgt acttatcgtc
      601 atagtagcag ataaaggggc taacggggga gggatcataa acctcacgtt cagggcactc
      661 atgcccgtag tggcccttct gtgcgcaatg gtaacaatat gccaacgctg aggggcgtga
      721 tggggttttg ggtttcttgg gggggcctgg cttagtagtc agatggtatt gacgccagat
      781 ttctgtgcaa gcgtcggtgt aatgaccaag catgtgacag cgatcacatt gtttgtccca
      841 gctatggcga aataaacagg agtggtccag catcttggga actggacagt actcgcaaag
      901 cggcgccgga cacgaataca gcaagtgccc acgacgggag caaaggaaac aacgacgcac
      961 tttacgcggc aatggacagt ttttagacag atgtccgcgt ttgtcacagt tgcggcaaat
     1021 gatattttta tttgcactat aataacgctg tgtccagcgt cccggtgtgc ggttatttgc
     1081 aatctgtgct tcgatatctt tatccgaaat cgaccaatta atcccatcct cgcgctccgg
     1141 tttgataggt tccgtggttg tgaagatatc tgaatagtgc tggcgctcaa atacgcgatc
     1201 aagaacctca agttgctgct gcgtaaaaag gtcaccacgc atttgcttac gcaaaaagtc
     1261 acgacctggc agcgagtgcc catttggaac cggtgactct tgaatcccct catcgcgctt
     1321 acgcttgttc gtatctgccg aaggcgatgt gatgcctaag atccccgaaa tcgagtaaga
     1381 ggaccccgca ctatctgtgc ttacagagct gacttgtgtg acactcccag tgctaacaat
     1441 actgtgggaa gaagccggaa cgggctggtt gggcggttgc tgaactttag tacgaataat
     1501 acgattaatg ctactgaccg atggaaccgt atcgttatca catacgcgtt cagccagtaa
     1561 gcggtcacgg atttcccagg caaacattgt cgggttctga cgcttatatt cggcgatctt
     1621 ctcgacgact ttcggggtgg ccacttttgg tttagaaccc ccaataacac ccggtttaat
     1681 actccccgtt tcatagtagc gccccaaaat cttagataca cagccgtgtg acacgcgtaa
     1741 ttggcggctg atatcacatg gacgtacgcc ttggtgcgcc aactctacaa tgcgctggcg
     1801 gaccacatca ggcaaagggc ggccattcac gaatacgcca ccaagctgat ttacacctcc
     1861 gtgcccggta cgtgaagtgc gaggtgtggg gtagttcttt tcaaggtcca ttccccggga
     1921 agcggccgca cttcccccgg agcctccgct tccgcccttg tacagctcgt ccatgccgag
     1981 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     2041 gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     2101 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     2161 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     2221 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     2281 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     2341 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     2401 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     2461 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     2521 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     2581 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     2641 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat cccgggcccg
     2701 cggtaccgtc gactgcagaa ttcggttcac taaacgagcc cggttcacta aacgagctct
     2761 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     2821 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     2881 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     2941 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     3001 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     3061 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     3121 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     3181 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     3241 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     3301 aattgattac tattaataac tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt
     3361 gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa
     3421 agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat gaataatttt
     3481 atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat
     3541 tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt
     3601 attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa
     3661 atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg
     3721 agttttccag agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca
     3781 gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt
     3841 tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt
     3901 ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag
     3961 aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc
     4021 tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat
     4081 tcacttctcc aattgtccct catatctcct ccttatggtg ttttactaaa cttttccatg
     4141 ttctaatcct catcctgtct acttgccaca caatcatcac ctgccatctg ttttccataa
     4201 tccctaatga tctttgcttt tcttcttggc actactttta tgtcactatt atcttgtatt
     4261 actactgccc cttcaccttt ccagaggagc tttgctggtc ctttccaaag tggatttctg
     4321 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     4381 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     4441 cccccttttc ttttaaaatt gtggatgaat actgccattt gtactgctgt cttaagatgt
     4501 tcagcctgat ctcttacctg tcctataatt ttctttaatt ctttattcat agattctact
     4561 actccttgac tttggggatt gtagggaatt tccaggtctg aagatcctgc tgcttgctgt
     4621 gcttttttct tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct
     4681 tttatctcta tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta
     4741 agttcttctg atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct
     4801 tctgatgttt ctaacaggcc agtattaact gcgaatcgtt ctagctccct gcttgcccat
     4861 actatatgtt ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat
     4921 cgcgatctaa ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag
     4981 cctccgctag tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg
     5041 cgcgcttcag caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc
     5101 aagtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     5161 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     5221 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     5281 tctggtctaa ccagagagac ccagtacagg caaaaagcag ctgcttatat gcaggatctg
     5341 agggctcgcc actccccagt cccgcccagg ccacgcctcc catggtaata gcgatgacta
     5401 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     5461 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     5521 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     5581 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     5641 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     5701 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     5761 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5821 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5881 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5941 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6001 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6061 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6121 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6181 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6241 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6301 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6361 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6421 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6481 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6541 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6601 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6661 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6721 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6781 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6841 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6901 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6961 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7021 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7081 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7141 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7201 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7261 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7321 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7381 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7441 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7501 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7561 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7621 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7681 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7741 cattaatgca gcggatccag acatgataag atacattgat gagtttggac aaaccacaac
     7801 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     7861 aaccattata agctgcaata aacaagttag ctttttgcaa aagcctaggc ctccaaaaaa
     7921 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     7981 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     8041 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgcg gatccgctgg
     8101 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     8161 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     8221 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
     8281 gggctgcagg tcgagggatc tccataagag aagagggaca gctatgactg ggagtagtca
     8341 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
     8401 aaaaaataac acaaaacaaa atat
//
