LOCUS       sequence-181972-        5391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of FtnA-Halo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  AUTHORS   Finan K, Raulf A, Heilemann M
  TITLE     A set of homo-oligomeric standards allows accurate protein counting.
  JOURNAL   Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12049-52. doi: 
            10.1002/anie.201505664. Epub 2015 Aug 20.
  PUBMED    26289028
REFERENCE   2  (bases 1 to 5391)
  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: 
            "10.1002/anie.201505664"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2015-10-5- 5"; volume: "54"; issue: "41"; pages: "12049-52"
COMMENT     Sequence Label: pFtnA-Halo
FEATURES             Location/Qualifiers
     source          1..5391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     502..521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        776..1079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1080..1283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1234..1254
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1322..2212
                     /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    2894..3015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2985..3004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3022..3477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3159..3180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3371..3390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3504..3608
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3610..3967
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3610..3801
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3633..3653)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3818..3953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4002..4796
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4002..4796
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4056..4075)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4666..4685
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4984..5003)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5028..5075
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
       61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
      721 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      781 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      841 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      901 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      961 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1021 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1081 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1141 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1201 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1261 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgccac cggtcgccac
     1321 catggcagaa atcggtactg gctttccatt cgacccccat tatgtggaag tcctgggcga
     1381 gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt tcctgcacgg
     1441 taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac cgacccatcg
     1501 ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc tgggttattt
     1561 cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc tggaagaggt
     1621 cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca agcgcaatcc
     1681 agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga cctgggacga
     1741 atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg tcggccgcaa
     1801 gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg tcgtccgccc
     1861 gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg ttgaccgcga
     1921 gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga acatcgtcgc
     1981 gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc tgctgttctg
     2041 gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca aaagcctgcc
     2101 taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag acaacccgga
     2161 cctgatcggc agcgagatcg cgcgctggct gtctactctg gagatttccg gcgctagcgg
     2221 agcatccgca ggaatgctga aaccagaaat gattgaaaaa cttaatgagc agatgaacct
     2281 ggaactgtac tcttcactgc tttatcagca aatgagcgcc tggtgcagct atcatacctt
     2341 cgaaggtgct gccgcgttcc tgcgccgtca cgcccaggaa gagatgacgc atatgcagcg
     2401 tctgtttgat tacctgactg ataccggcaa tttaccgcgt attaataccg ttgaatctcc
     2461 gtttgctgaa tattcctcac ttgatgaatt attccaggaa acctataaac acgaacaatt
     2521 aatcacccag aaaattaacg aactggctca tgctgcaatg accaatcagg actacccaac
     2581 atttaatttc ctgcaatggt atgtttctga gcagcatgaa gaagagaaac tgttcaaatc
     2641 gattattgat aaattaagcc tggcaggcaa aagcggcgaa ggtctgtatt ttatcgacaa
     2701 agaactctct accctcgaca cacaaaacta acggccgcat cgataacacc gcaaatgctt
     2761 ctagggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     2821 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2881 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3001 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3061 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3121 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3181 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3241 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3301 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3361 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3421 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3541 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3601 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3841 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3901 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3961 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4501 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4621 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4801 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4861 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4921 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4981 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5041 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5101 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5161 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5221 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5281 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5341 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga t
//
