LOCUS       sequence-338322-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of hH2A-(GGS)X6-HaloTag C58 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Shao S, Zhang H, Zeng Y, Li Y, Sun C, Sun Y
  TITLE     TagBiFC technique allows long-term single-molecule tracking of 
            protein-protein interactions in living cells.
  JOURNAL   Commun Biol. 2021 Mar 19;4(1):378. doi: 10.1038/s42003-021-01896-7.
  PUBMED    33742089
REFERENCE   2  (bases 1 to 6536)
  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: "Commun 
            Biol."; date: "2021-03-19"; volume: "4(1):378. doi"; pages: " 
            10.1038/s42003-021-01896-7"
COMMENT     Sequence Label: pCDNA3.1(+)-hH2A-(GGS)X6-HaloTag C58
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2127..2144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2133..2357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2403..2831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2490..2509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2700..2721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2840..2860)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2845..3174
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3025..3160
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3087..3106
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3241..4035
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3295..3314)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3905..3924
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4209..4330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4246..4265)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4300..4319
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4379..4395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4379..4395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4392..4414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4403..4419
                     /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)."
     promoter        complement(join(4427..4433,4434..4451,4452..4457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4472..4493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4610..4627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4781..5369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4861..4880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5540..6331,6332..6400))
                     /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     6163..6182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6401..6505)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgt ctggacgtgg caagcagggc ggcaaggctc gcgccaaggc
      961 caaaacccgc tcctctagag ctgggctcca atttcctgta ggacgagtgc accgcctgct
     1021 ccgcaagggc aactacgctg agcgggtcgg ggccggcgcg ccggtttacc tggcggcggt
     1081 gctggagtac ctaactgccg agatcctgga gctggcgggc aacgcagccc gcgacaacaa
     1141 aaagacccgc atcatcccgc gccacttgca gctggccatc cgcaacgacg aggagctcaa
     1201 caagctgctt ggtaaagtta ccatcgctca gggcggtgtt ctgcctaaca tccaggccgt
     1261 actgctcccc aagaagactg agagccacca caaagctaag ggcaagggat ccggcggttc
     1321 tggaggctcc ggcggttcag ggggatcggg cggctcaggc ggctctgaat tcacccaccg
     1381 ttgtattgca ccggacctga tcggtatggg caagagtgac aagccggacc tgggttattt
     1441 cttcgacgac cacgtgcgct tcatggatgc attcatcgag gccctgggcc tggaagaggt
     1501 ggttctggtg atccatgatt ggggcagcgc cctgggcttt cattgggcca agcgcaaccc
     1561 ggagcgtgtg aagggcatcg ccttcatgga gttcatccgc cctatcccga cctgggatga
     1621 gtggccggaa tttgcccgcg agacctttca ggccttccgt accacagacg ttggccgcaa
     1681 gctgatcatc gatcagaacg tgtttatcga gggcaccctg cctatgggtg tggttcgccc
     1741 tctgaccgag gtggagatgg accactaccg cgagccgttc ctgaatccgg ttgatcgcga
     1801 gccgctgtgg cgcttcccga atgagctgcc gatcgccggc gaacctgcca atatcgtggc
     1861 cctggtggag gagtacatgg actggttaca ccagagcccg gtgcctaagc tgctgttctg
     1921 gggtaccccg ggcgtgctga ttccgcctgc agaagccgcc cgcctggcaa agagtctgcc
     1981 gaattgcaag gccgtggaca tcggtccggg tctgaatctg ctgcaagagg acaacccgga
     2041 tctgatcggt agtgagatcg cccgctggct gagtacactg gagattagcg gctaatctag
     2101 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2161 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2221 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2281 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2341 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2401 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2461 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2581 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2701 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2761 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2821 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2881 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2941 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3001 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3121 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3181 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3241 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3301 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3361 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3421 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3481 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3541 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3601 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3661 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3721 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3781 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3841 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3901 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3961 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4021 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4081 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4141 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4201 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4261 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4321 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4381 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4441 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4501 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4561 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4621 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4681 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
