LOCUS       Exported                5741 bp ds-DNA     circular SYN 25-11-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    SNAPtag-PixE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5741)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported 2025¤¤¤Å¤¤ 11 25¤·¤Ó¤© ¤¾¤È¤·¤Ë¤·¤Ó¤© from SnapGene 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5741
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             627..1172
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /note="SNAP-tag(R)"
                     /note="optimized for mammalian expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(1207..1224)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_feature    1251
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1257..2399
                     /codon_start=1
                     /note="PixE"
                     /translation="MSNSVLSTSSPGNIQTRQAIPAQLLYSLVNQKASGKLVVQNPFDE
                     FVYWQVYLGDGRIHLANSANGCSERLNYLIGSTLRQKGIELPSRIADDYIFLSGLWKRD
                     VFSFQQTRSILTQASQEALVQVLSLPKTSYRFNQEKELSELFLNLELEKTITPIKHKIR
                     YWWELRSYINSPFQRPLVENWLSLRQNLVKNNLHGQHWLKRFHTCLENLNCLYEIAGHL
                     EISTLELALMLQPMVKTGEIKMLPYQEIQTDDRPLVAYVDHREVQQRMMTYSLEKEGYR
                     ALVLDDPFRALTVLLNSPPNLIIINTDLPDMNGYQLCSFCRKSNQLKDVPILLLGKGDN
                     LISGIRIKLCQASGYLSEPFVPEELLDRVAKLLPTNKTNS"
     misc_feature    2405
                     /note="MCS"
                     /note="multiple cloning site"
     polyA_signal    2529..2650
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2657..3112)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3139..3243
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        3245..3602
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3453..3588
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3637..4431
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4663..4710
                     /note="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal"
     rep_origin      5039..5627
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atccaccggt gccaccatgg acaaagactg cgaaatgaag cgcaccaccc
      661 tggatagccc tctgggcaag ctggaactgt ctgggtgcga acagggcctg cacgagatca
      721 agctgctggg caaaggaaca tctgccgccg acgccgtgga agtgcctgcc ccagccgccg
      781 tgctgggcgg accagagcca ctgatgcagg ccaccgcctg gctcaacgcc tactttcacc
      841 agcctgaggc catcgaggag ttccctgtgc cagccctgca ccacccagtg ttccagcagg
      901 agagctttac ccgccaggtg ctgtggaaac tgctgaaagt ggtgaagttc ggagaggtca
      961 tcagctacca gcagctggcc gccctggccg gcaatcccgc cgccaccgcc gccgtgaaaa
     1021 ccgccctgag cggaaatccc gtgcccattc tgatcccctg ccaccgggtg gtgtctagct
     1081 ctggcgccgt ggggggctac gagggcgggc tcgccgtgaa agagtggctg ctggcccacg
     1141 agggccacag actgggcaag cctgggctgg gtatgtacag tgggggaggc ggagggggtg
     1201 gttcaggtgg tggtggtggt ggtggttcgg gtggtggcgg agggggaggc tccggaatga
     1261 gcaattcagt tttgtccact tcttccccag gtaatatcca aactcggcag gcgattccgg
     1321 cccaactcct ctattccctc gtcaaccaaa aagccagcgg caaacttgtg gtacaaaacc
     1381 cctttgacga atttgtttat tggcaagttt atttggggga tggtcgtatt cacttggcca
     1441 acagcgccaa tggttgttcc gaaagactta attatctgat tggtagcacc cttcgtcaga
     1501 agggcattga actgccatca agaattgctg acgattatat ttttctttcc gggctctgga
     1561 aaagggatgt tttttccttt cagcaaaccc gctcgattct gacccaagct agccaagaag
     1621 ccttagttca ggtactttcc ttacctaaaa catcctatcg ttttaatcaa gaaaaagaac
     1681 tgagcgaact atttttgaat ttagagttag aaaaaactat tacccccatc aaacataaaa
     1741 ttcgctattg gtgggaattg aggtcctata ttaactcgcc tttccagcga cccctagtgg
     1801 aaaactggct tagtctgcgc caaaatttgg tgaaaaataa cctccatggc caacattggc
     1861 taaaacgctt ccacacttgc ttggaaaatc taaattgctt atatgaaatt gccgggcatt
     1921 tggaaatcag tactttggag ttggcattaa tgctacagcc catggtcaag actggcgaaa
     1981 tcaagatgct gccttatcaa gaaattcaaa cggatgaccg ccctttggtg gcttatgttg
     2041 accataggga agtccagcag cgcatgatga cctattcttt ggaaaaagaa ggctatcgag
     2101 ccttagtact ggatgatccc tttcgggctt taacggtgtt gttgaattct ccccccaatc
     2161 tgattatcat aaatactgac ttgccggaca tgaacggtta tcaactctgt tcgttttgcc
     2221 gtaagtccaa tcaacttaaa gatgtgccga tcctactatt ggggaaaggt gacaatttga
     2281 tcagcggtat tcgtatcaaa ctttgtcaag cttctgggta cttaagtgag ccttttgttc
     2341 cagaagaatt actcgatcgg gtggcaaaac tccttcccac caataaaacc aactcctgag
     2401 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2461 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2521 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2641 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2701 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2761 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2821 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2881 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2941 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3001 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3061 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3241 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3541 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3601 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3661 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3721 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3781 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3841 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3901 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3961 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4021 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4081 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4141 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4201 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4261 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4321 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4381 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4441 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4501 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4561 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4621 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4681 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4741 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4801 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4861 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4921 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5701 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
