LOCUS       sequence-388534-        5995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein production of the split HaloTag part cpHalo∆ 
            (cpHaloTag135/156) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5995)
  AUTHORS   Huppertz MC, Wilhelm J, Grenier V, Schneider MW, Falt T, Porzberg N,
            Hausmann D, Hoffmann DC, Hai L, Tarnawski M, Pino G, Slanchev K, 
            Kolb I, Acuna C, Fenk LM, Baier H, Hiblot J, Johnsson K
  TITLE     Recording physiological history of cells with chemical labeling.
  JOURNAL   Science. 2024 Feb 23;383(6685):890-897. doi: 
            10.1126/science.adg0812. Epub 2024 Feb 22.
  PUBMED    38386755
REFERENCE   2  (bases 1 to 5995)
  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"; doi: 
            "10.1126/science.adg0812"; journalName: "Science"; date: 
            "2024-02-23- 23"; volume: "383"; issue: "6685"; pages: "890-897"
COMMENT     Sequence Label: pET-51b(+)-cpHalo∆
FEATURES             Location/Qualifiers
     source          1..5995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(155..1237)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1199..1218
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1238..1315)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1521..1540
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1628..1647
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1628..1646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1647..1671
                     /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)."
     RBS             1686..1708
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1718..1747
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1748..1768
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2670..2688)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2684..2731
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2768..3223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2855..2874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3065..3086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3338..4129,4130..4198))
                     /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     3961..3980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4199..4291)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4372..4960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4861..4880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5114..5131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5146..5285
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5211..5230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5371..5393
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5387..5578)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccagcctagc cgggtcctca acgacaggag cacgatcatg ctagtcatgc cccgcgccca
       61 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
      121 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      181 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      241 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
      301 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
      361 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
      421 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
      481 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
      541 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
      601 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
      661 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
      721 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
      781 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
      841 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
      901 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
      961 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     1021 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     1081 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     1141 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     1201 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     1261 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     1321 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     1381 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     1441 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     1501 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     1561 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc
     1621 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     1681 ataattttgt ttaactttaa gaaggagata taccatgcat caccaccatc atcaccatca
     1741 ccaccacgaa aacctgtatt ttcagggcga cgtcggccgc aagctgatca tcgatcagaa
     1801 cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat
     1861 ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc
     1921 aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat
     1981 ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct
     2041 gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga
     2101 catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat
     2161 cgcgcgctgg ctgtctactc tggagattgg tggcaccggt ggctctggcg gtaccggtgg
     2221 ctctggcggt tctatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg
     2281 cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca
     2341 cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca
     2401 tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta
     2461 tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga
     2521 ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa
     2581 tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga
     2641 cgaatggtaa ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2701 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     2761 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2821 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2881 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2941 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3001 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3061 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3121 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3181 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttctggcgg cacgatggca
     3241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3421 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3481 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3541 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3601 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3661 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3721 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3781 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3841 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3901 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3961 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4021 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4081 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4141 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4201 tcttcctttt tcaatcatga ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4261 atatttgaat gtatttagaa aaataaacaa ataggtcatg accaaaatcc cttaacgtga
     4321 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4381 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4441 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4501 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4561 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4621 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4681 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4741 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4801 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4861 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4921 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4981 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5041 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5101 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     5161 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     5221 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     5281 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5341 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5401 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     5461 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     5521 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     5581 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     5641 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     5701 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     5761 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     5821 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     5881 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     5941 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccg
//
