LOCUS       sequence-427418-        4900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression G4PID-HA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4900)
  AUTHORS   Lu Z, Xie S, Su H, Han S, Huang H, Zhou X
  TITLE     Identification of G-quadruplex-interacting proteins in living cells 
            using an artificial G4-targeting biotin ligase.
  JOURNAL   Nucleic Acids Res. 2024 Mar 7:gkae126. doi: 10.1093/nar/gkae126.
  PUBMED    38452210
REFERENCE   2  (bases 1 to 4900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2024 Mar 7:gkae126. doi: 10.1093/nar/gkae126."
COMMENT     Sequence Label: p-G4PID-HA
FEATURES             Location/Qualifiers
     source          1..4900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             604..630
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             769..1539
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     polyA_signal    1688..1809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1725..1744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1779..1798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1816..2271)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1953..1974)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2165..2184
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2298..2402
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2404..2761
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2404..2595
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2427..2447)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2612..2747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2796..3590
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2796..3590
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2850..2869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3460..3479
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3778..3797)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3822..3869
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4198..4786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4687..4706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
      601 atgtacccat acgatgttcc agattacgct agatctgcca ccatgcatcc tggccatctc
      661 aaaggccgtg aaatcggcat gtggtacgct aagaaacagg gtcagaaaaa caaaggcacg
      721 ggaagcggcg caggcaccgg cagcggtgcg ggtaccggct ccggcgcaat cccgctgctg
      781 aacgctaaac agattctggg acagctggac ggcgggagcg tggcagtcct gcctgtggtc
      841 gactccacca atcagtacct gctggatcga atcggcgagc tgaagagtgg ggatgcttgc
      901 attgcagaat atcagcaggc agggagagga agcagaggga ggaaatggtt ctctcctttt
      961 ggagctaacc tgtacctgag tatgttttgg cgcctgaagc ggggaccagc agcaatcggc
     1021 ctgggcccgg tcatcggaat tgtcatggca gaagcgctgc gaaagctggg agcagacaag
     1081 gtgcgagtca aatggcccaa tgacctgtat ctgcaggata gaaagctggc aggcatcctg
     1141 gtggagctgg ccggaataac aggcgatgct gcacagatcg tcattggcgc cgggattaac
     1201 gtggctatga ggcgcgtgga ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa
     1261 gcagggatta acctggacag gaatactctg gccgctatgc tgatccgaga gctgcgggca
     1321 gccctggaac tgttcgagca ggaaggcctg gctccatatc tgtcacggtg ggagaagctg
     1381 gataacttca tcaatagacc cgtgaagctg atcattgggg acaaagagat tttcgggatt
     1441 agccggggga ttgataaaca gggagccctg ctgctggaac aggacggagt tatcaaaccc
     1501 tggatgggcg gagaaatcag tctgcggtct gccgaaaagt aagaattctg caggtcgagc
     1561 atgcatgcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     1621 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1681 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1741 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1801 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1861 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     1921 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     1981 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2041 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2101 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2161 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2221 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2281 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2341 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2401 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2461 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2521 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2581 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2641 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2701 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2761 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2821 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2881 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2941 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3001 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3061 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3121 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3181 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3241 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3301 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3361 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3421 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3481 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3541 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3601 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3661 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3721 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     3781 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3841 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3901 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3961 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4021 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4081 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4141 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4201 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4261 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4321 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4381 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4441 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4501 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4561 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4621 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4681 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4741 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4801 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4861 atcccctgat tctgtggata accgtattac cgccatgcat
//
