LOCUS       sequence-386422-        8238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  single-particle tracking for H2AZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8238)
  AUTHORS   Dong P, Zhang S, Xie L, Wang L, Lemire AL, Lander AD, Chang HY, Liu 
            ZJ
  TITLE     Spatial organization of the 3D genome encodes gene co-expression 
            programs in single cells
  JOURNAL   bioRxiv 2022.10.26.513917
REFERENCE   2  (bases 1 to 8238)
  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: "bioRxiv 
            2022.10.26.513917"
COMMENT     Sequence Label: piggybac-EF1-H2AZ-halo
FEATURES             Location/Qualifiers
     source          1..8238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        135..346
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             359..627
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1013..1033
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1049..1933
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     misc_feature    1968..2540
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2135..2152)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2362..2381
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2549..3343
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2603..2622)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3213..3232
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3403..3423)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3465..3586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3556..3575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4349..4383
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4874..4890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4874..4890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4887..4909)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4898..4914
                     /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(4922..4928,4929..4946,4947..4952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4967..4988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5105..5122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5276..5864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5356..5375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6035..6826,6827..6895))
                     /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     6658..6677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6896..7000)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7026..7481)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7163..7184)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7375..7394
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7608..7630
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7622..7639
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7623..7639
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7769..7831
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ggcctccacg gccaattcca tatgccaagg atctgcgatc gctccggtgc ccgtcagtgg
       61 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattcgctc
      121 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
      181 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
      241 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
      301 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
      361 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
      421 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
      481 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
      541 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
      601 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg acgcgtatgg ctggcggtaa
      661 ggctggaaag gactccggaa aggccaagac aaaggcggtt tcccgctcgc agcgagccgg
      721 cttgcagttc cctgtgggcc gtattcatcg acacctgaaa tctaggacaa ccagccacgg
      781 acgtgtgggc gcgaccgccg ctgtgtacag cgcagccatc ctggagtacc tcaccgcaga
      841 gcttgctata cgtggagatg aagaattgga ttctctgatc aaagctacca ttgctggtgg
      901 tggtgtcatc ccacacatcc acaaatcgct gatcgggaag aaaggacaac agaagactgt
      961 ttccggtggc ggcggatctg gcggtggtgg gtccggagga ggcggttcga tggaggatct
     1021 gtactttcag agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca
     1081 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac
     1141 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc
     1201 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga
     1261 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga
     1321 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt
     1381 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg
     1441 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg
     1501 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct
     1561 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt
     1621 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg
     1681 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc
     1741 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc
     1801 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct
     1861 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct
     1921 ggagatttcc ggttaagaat tcggcttctc gagggattgc ggccgcgccc ctctccctcc
     1981 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2041 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2101 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2161 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2221 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2281 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2341 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2401 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     2461 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2521 ctttgaaaaa cacgataata cctccggaat gattgaacaa gatggattgc acgcaggttc
     2581 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2641 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2701 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     2761 cgcgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2821 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2881 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2941 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3001 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3061 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3121 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3181 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3241 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3301 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagtcgaca atcaacctct
     3361 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3421 atgtggatac gctgctttaa tgcctttgta tcatgcgtta actaaacttg tttattgcag
     3481 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3541 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggaatt
     3601 gactcaaatg atgtcaatta gtctatcaga agctcatctg gtctcccttc cgggggacaa
     3661 gacatccctg tttaatattt aaacagcagt gttcccaaac tgggttctta tatcccttgc
     3721 tctggtcaac caggttgcag ggtttcctgt cctcacagga acgaagtccc taaagaaaca
     3781 gtggcagcca ggtttagccc cggaattgac tggattcctt ttttagggcc cattggtatg
     3841 gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag
     3901 aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc
     3961 ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct
     4021 agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt ccctgatatc
     4081 tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata
     4141 taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga
     4201 ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca
     4261 cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt
     4321 agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta
     4381 gggttaatct agctgcatca ggatcatatc gtcgggtctt ttttccggct cagtcatcgc
     4441 ccaagctggc gctatctggg catcggggag gaagaagccc gtgccttttc ccgcgaggtt
     4501 gaagcggcat ggaaagagtt tgccgaggat gactgctgct gcattgacgt tgagcgaaaa
     4561 cgcacgttta ccatgatgat tcgggaaggt gtggccatgc acgcctttaa cggtgaactg
     4621 ttcgttcagg ccacctggga taccagttcg tcgcggcttt tccggacaca gttccggatg
     4681 gtcagcccga agcgcatcag caacccgaac aataccggcg acagccggaa ctgccgtgcc
     4741 ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt acgtcagcga ggacgggtat
     4801 cctggctgga tgccgcagaa atggacatgg ataccccgtg agttacccgg cgggcgcgct
     4861 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4921 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4981 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5041 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5101 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5161 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5221 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5281 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5341 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5401 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5461 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5521 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5581 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5641 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5701 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5761 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5821 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5881 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5941 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6001 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6061 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6121 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6181 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6241 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6301 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6361 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6421 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6481 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6541 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6601 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6661 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6721 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6781 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6841 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6901 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6961 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7021 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     7081 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     7141 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     7201 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7261 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7321 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7381 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7441 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     7501 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     7561 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7621 ttgtaaaacg acggccagtg agcgcgcctc gttcattcac gtttttgaac ccgtggagga
     7681 cgggcagact cgcggtgcaa atgtgtttta cagcgtgatg gagcagatga agatgctcga
     7741 cacgctgcag aacacgcagc tagattaacc ctagaaagat aatcatattg tgacgtacgt
     7801 taaagataat catgtgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta
     7861 tttattaatt tgaatagata ttaagtttta ttatatttac acttacatac taataataaa
     7921 ttcaacaaac aatttattta tgtttattta tttattaaaa aaaacaaaaa ctcaaaattt
     7981 cttctataaa gtaacaaaac ttttatgagg gacagccccc ccccaaagcc cccagggatg
     8041 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
     8101 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
     8161 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
     8221 ggggggatac ggggaaaa
//
