LOCUS       sequence-338443-        5990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HaloTag N58-(GGS)X6-hH2A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  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 5990)
  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(+)-HaloTag N58-(GGS)X6-hH2A
FEATURES             Location/Qualifiers
     source          1..5990
                     /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(1581..1598)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1587..1811
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1857..2285
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1944..1963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2154..2175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2294..2314)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2299..2628
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2479..2614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2541..2560
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2695..3489
                     /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(2749..2768)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3359..3378
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3663..3784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3700..3719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3754..3773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3833..3849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3833..3849)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3846..3868)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3857..3873
                     /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(3881..3887,3888..3905,3906..3911))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3926..3947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4064..4081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4235..4823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4315..4334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4994..5785,5786..5854))
                     /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     5617..5636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5855..5959)
                     /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 aagcttatgg gcagcgaaat tggcacaggc ttcccgttcg acccgcatta
      961 cgttgaggtg ctgggcgagc gcatgcacta tgtggacgtg ggtccgcgtg acggtacccc
     1021 tgtgctgttc ctgcatggta acccgaccag cagctacgtg tggcgcaaca ttatcccgca
     1081 cgtggccccg ggatccggcg gttctggagg ctccggcggt tcagggggat cgggcggctc
     1141 aggcggctct gaattcatgt ctggacgtgg caagcagggc ggcaaggctc gcgccaaggc
     1201 caaaacccgc tcctctagag ctgggctcca atttcctgta ggacgagtgc accgcctgct
     1261 ccgcaagggc aactacgctg agcgggtcgg ggccggcgcg ccggtttacc tggcggcggt
     1321 gctggagtac ctaactgccg agatcctgga gctggcgggc aacgcagccc gcgacaacaa
     1381 aaagacccgc atcatcccgc gccacttgca gctggccatc cgcaacgacg aggagctcaa
     1441 caagctgctt ggtaaagtta ccatcgctca gggcggtgtt ctgcctaaca tccaggccgt
     1501 actgctcccc aagaagactg agagccacca caaagctaag ggcaagtaat ctagagggcc
     1561 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1621 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1681 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1741 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1801 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1861 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1921 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1981 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2041 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2101 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2161 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2221 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2281 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2401 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2461 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2581 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2641 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2701 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2761 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2821 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2881 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2941 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3001 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3061 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3121 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3181 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3241 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3301 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3361 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3421 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3481 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3541 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3601 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3661 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3721 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3781 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3841 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3901 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3961 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4021 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4081 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4861 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4921 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4981 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5041 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5101 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5161 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5221 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5281 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5341 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5401 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5461 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5521 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5581 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5641 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5701 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5761 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5821 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5881 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5941 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
