LOCUS       sequence-463625-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lac repressor (LacI) which binds lacO sites; labeled with HaloTag 
            and coupled with effector protein mouse HP1alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  TITLE     HP1 binding creates a local barrier against transcription activation
            and persists during chromatin decondensation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7793)
  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: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-LacI-mHP1a
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2028..2047)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3744..3762)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3794..3915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3805..3824)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3859..3878
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4160..4517
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4160..4351
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4183..4203)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4368..4503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4568..5362
                     /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(4622..4641)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5232..5251
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5426..5474
                     /note="synthetic polyadenylation signal"
     promoter        5630..5734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5735..5803,5804..6595)
                     /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(5953..5972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6753..7341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6833..6852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7457..7740
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccaat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgaaatggt gaaaccagta acgttatacg atgtcgcaga
     2041 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     2101 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     2161 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     2221 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     2281 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     2341 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     2401 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     2461 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     2521 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     2581 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     2641 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     2701 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     2761 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     2821 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     2881 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     2941 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     3001 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3061 acaggtttcc cgaagcagcc tggagctgta caagtccgga atgggaaaga agaccaagag
     3121 gacagccgac agctcttctt cagaggatga ggaggaatat gtggtggaaa aggtgttgga
     3181 caggcgcatg gttaaggggc aagtggaata tctgttgaag tggaaaggct tttctgagga
     3241 gcacaatact tgggaacctg agaagaactt ggattgtcct gaactaattt ctgagtttat
     3301 gaaaaagtat aagaagatga aggagggtga aaacaataag cccagggaga aatcagaagg
     3361 aaacaagagg aaatccagtt tctccaacag cgctgatgat attaaatcta aaaaaaagag
     3421 agagcaaagc aatgatatcg ctcggggctt tgagagagga ctggaaccag aaaagatcat
     3481 cggagcaaca gattcctgcg gtgacttaat gttcttaatg aaatggaaag acacagatga
     3541 agctgacctg gttcttgcaa aagaagctaa cgtgaagtgt ccacagattg tgatagcatt
     3601 ttatgaagag agactgacgt ggcacgcata tccagaggat gcggaaaaca aagaaaaaga
     3661 aagcgcgaag agctaaggat ccaccggatc tagataactg atcataatca gtgagcggcc
     3721 gcttgctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggccgctt
     3781 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3841 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3901 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3961 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
     4021 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     4081 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     4141 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4441 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4501 gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag
     4561 agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4741 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5341 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5401 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5461 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     5521 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     5581 ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5641 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5701 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5761 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5821 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5881 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5941 tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6001 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6061 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6121 taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt
     6181 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6241 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6301 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6361 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6421 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6481 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6541 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga
     6601 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta
     6661 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6721 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6781 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6841 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6901 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6961 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7021 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7081 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7141 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7201 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7261 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7321 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7381 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7441 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     7501 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     7561 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     7621 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     7681 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     7741 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
