LOCUS       sequence-362159-        4998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector backbone to clone insert downstream of Halo tag (Halo tag 
            codon optimized for Xenopus), for mRNA synthesis .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4998)
  AUTHORS   Dudley CE, van den Goor L, Miller AL
  TITLE     SNAP- and Halo-tagging and dye introduction protocol for live 
            microscopy in Xenopus embryos.
  JOURNAL   STAR Protoc. 2022 Aug 18;3(3):101622. doi: 
            10.1016/j.xpro.2022.101622. eCollection 2022 Sep 16.
  PUBMED    36035797
REFERENCE   2  (bases 1 to 4998)
  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: "STAR 
            Protoc."; date: "2022-08-18"; volume: "3(3):101622. doi"; pages: " 
            10.1016/j.xpro.2022.101622. eCollection 2022 Sep 16"
COMMENT     Sequence Label: pCS2+/N-Halo
FEATURES             Location/Qualifiers
     source          1..4998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        355..373
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     355..372
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             410..1300
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    1370..1504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1394..1413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1448..1467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1617..1637)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1617..1635)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1656..1672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1656..1672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1669..1691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1680..1696
                     /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(1704..1710,1711..1728,1729..1734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1749..1770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1887..1904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2058..2646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2138..2157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2817..3608,3609..3677))
                     /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     3440..3459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3678..3782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3808..4263)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3945..3966)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4157..4176
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4334..4998,1..320)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
       61 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
      121 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
      181 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
      241 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
      301 gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag cttgatttag
      361 gtgacactat agaatacaag ctacttgttc tttttgcagg atccaaacca tggcagaaat
      421 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
      481 cgtcgatgtt ggtccacgcg atggcacccc tgtgctgttc ctgcacggta acccaacctc
      541 ctcctacgtg tggcgcaaca tcatcccaca tgttgcacca acccatcgct gcattgctcc
      601 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
      661 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
      721 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
      781 aggtattgca tttatggagt tcatccgccc tatcccaacc tgggacgaat ggccagaatt
      841 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
      901 tcagaacgtt tttatcgagg gtacactgcc aatgggtgtc gtccgcccac tgactgaagt
      961 cgagatggac cattaccgcg agccattcct gaatcctgtt gaccgcgagc cactgtggcg
     1021 cttcccaaac gagctgccaa tcgccggtga gccagctaac atcgtcgctc tggtcgaaga
     1081 atacatggac tggctgcacc agtcccctgt cccaaagctg ctgttctggg gcaccccagg
     1141 cgttctgatc ccaccagccg aagccgctcg cctggccaaa agcctgccta actgcaaggc
     1201 tgtggacatc ggcccaggtc tgaatctgct gcaagaagac aacccagacc tgatcggcag
     1261 cgagatcgct cgctggctga gcacactcga gatttccggc tccggactca gatctcgagc
     1321 tcaagcttcg aattctgcag tctagaacta tagtgagtcg tattacgtag atccagacat
     1381 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1501 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1561 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     1621 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1681 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     1741 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1801 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1861 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1921 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1981 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2221 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2461 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2881 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2941 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3001 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3061 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3781 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     3841 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3901 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3961 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4021 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4081 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4141 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4201 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4261 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4321 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     4381 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4441 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4501 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     4561 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     4621 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     4681 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     4741 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     4801 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     4861 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     4921 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     4981 ccacccacca ttgacgtc
//
