LOCUS       sequence-194162-        3725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of protein of interest with SNAP-Tag in bacterial cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3725)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3725)
  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            "Unpublished"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: pSNAP-tag (T7) Vector
FEATURES             Location/Qualifiers
     source          1..3725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(535..540,541..558,559..565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    573..589
                     /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)."
     primer_bind     578..600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     597..613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     597..613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     632..652
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        634..652
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(737..784)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     780..798
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(919..1464)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     RBS             1472..1494
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1544..1563)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1545..1563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1573..1590)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1573..1589)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1582..1604)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1731..2186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1818..1837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2028..2049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2212..2316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2317..2385,2386..3177)
                     /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(2535..2554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3348..3725,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      541 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      601 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
      661 gctggagctc caccgcggtg gcggccgctc tagaactagt gcctctagat tatccggata
      721 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
      781 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
      841 ccggatctgc tcgagcggcc gccagtgtga tggatatctg cagaattctt aggatcctgg
      901 cgcgcctata cctgcaggtc ccagacccgg tttacccaga cgatgacctt catgggccag
      961 cagccactct ttcaccgcca gaccaccttc ataaccgcca accgcaccgc tagagctaac
     1021 cacacgatgg cacggaatca gaatcggcac cgggttaccg ctcagcgcgg ttttaacggc
     1081 ggcggtggcc gccggattac ccgccagggc cgccagctgc tgatagctaa tcacttcgcc
     1141 aaatttaacc actttcagca gtttccacag cacctgacgg gtaaagctct cctgctgaaa
     1201 caccggatga tgcagcgccg gaaccggaaa ttcttcaatc gcttccggct gatgaaaata
     1261 cgcgttcagc cacgcggtcg cctgcatcag cggttccgga ccacccagca cggcggccgg
     1321 ggccggaact tcaaccgcat cggccgcgct ggtgccttta cccagcagtt taatttcatg
     1381 caggccctgt tcgcagccgc tcagttccag tttgcccagc gggctatcca gggtggtacg
     1441 tttcatttcg caatctttgt ccatatgtat atctccttct taaagttaaa caaaattatt
     1501 tctagaggga aaccgttgtg gtctgggccc ggtacccaat tcgccctata gtgagtcgta
     1561 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1621 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1681 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     1741 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1801 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1861 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1921 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1981 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2041 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2101 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2161 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     2221 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2281 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2341 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2401 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2461 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2521 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2581 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2641 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2701 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2761 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2821 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2881 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2941 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3001 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3061 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3121 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3181 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3241 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3301 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3361 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3421 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3481 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3541 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3601 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3661 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3721 agata
//
