LOCUS       sequence-481956-        5401 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express HaloTag-SUMO1 Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5401)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5401)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-SUMO1 Vector
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    98..179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        325..535
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        347..543
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      394..529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      568..577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             574..972
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    996..1044
                     /note="synthetic polyadenylation signal"
     rep_origin      1170..1758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1659..1678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1916..2710)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2027..2046)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2637..2656
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2742..2789)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2799..2832
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2947..3068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3038..3057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3490..3510)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(3523..4413)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     regulatory      4410..4419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4428..4447)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4429..4447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4489..4508)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4491..4623
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(4759..4962)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4788..4808)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4963..5266
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
       61 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
      121 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgcc
      181 gacgtcttaa ttaacgcgtc ttgcgcagca ccatggcctg aaataacctc tgaaagagga
      241 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
      301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      361 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      481 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      541 aaaaagctcg attcttctga cactagcgcc accatggcca agcctttgtc tcaagaagaa
      601 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
      661 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
      721 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
      781 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
      841 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
      901 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
      961 tgggagggct aaactctagc catcacgatg gccgcaataa aatatcttta ttttcattac
     1021 atctgtgtgt tggttttttg tgtgacgatt ccggactata aaaggatcta ggtgaagatc
     1081 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1141 gaccccgtag aaaagatcaa aggatcttct tgaaatcctt tttttctgcg cgtaatctgc
     1201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1261 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     1321 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1381 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1441 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1501 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1561 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1621 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1681 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1741 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1801 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1861 accgcctttg agtgagctga taccggttgg gcgtcgcttg gtcggtcatt tcgaatcaga
     1921 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     1981 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     2041 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     2101 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     2161 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     2221 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     2281 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     2341 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     2401 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     2461 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     2521 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     2581 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     2641 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     2701 gttcaatcat gatctgcctc ctgggtttag ttcctcacct tgtcgtatta tactatgccg
     2761 atatactatg ccgatgatta attgtcaaca gtggatccat aacttcgtat aatgtatgct
     2821 atacgaagtt atattagaat tcttaattaa gattttacca catttgtaga ggttttactt
     2881 gcttaaaaaa acctcccaca tctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2941 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3001 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3061 gtatcttatc atgtctgctc gaagcggcct tgctcagcgg tggcagcagc caaatcagct
     3121 tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
     3181 ctaacccccc gtttgttcct gataaacttc aatcacatct tcttcctcca ttcccagttc
     3241 ttttggagta tgattatcag caattctctg accctcaaag agaaacctga gtgaattcat
     3301 tggaacaccc tgtctttgac agtatgattc tttgagtttc ttgagatgtg ttgtcatttt
     3361 cactttgaag tgaatctcac tgctatcctg tccaatgact ttgagtttaa tatattcacc
     3421 ttccttctta tcccccaagt cctcagttga aggttttgcc tcctggtcag acatggcgat
     3481 cgcgttatcg ctctgaaagt acagatcctc agtggttggc tcgccggaaa tctcgagcgt
     3541 cgacagccag cgcgcgatct cgctgccgat caggtccggg ttgtcttctt gcagcagatt
     3601 cagacccggg ccgatgtcca cagccttgca gttaggcagg cttttggcca ggcgagcggc
     3661 ttcggccggt gggatcagaa cgcctggggt gccccagaac agcagcttcg ggacagggga
     3721 ctggtgcagc cagtccatgt attcttcgac cagcgcgacg atgttcgctg gctcaccggc
     3781 gattggcagc tcgtttggga agcgccacag tggctcgcgg tcaacaggat tcaggaacgg
     3841 ctcgcggtaa tggtccatct cgacttcagt cagcgggcgg acgacaccca tcggcagcgt
     3901 accctcgata aaaacgttct gatcgatgat cagcttgcgg ccgacgtcgg tggtgcggaa
     3961 ggcctggaag gtctcgcggg caaattctgg ccattcgtcc caggtcggga tagggcggat
     4021 gaactccata aatgcaatac ctttgacgcg ctctggattg cgcttggccc agtggaaacc
     4081 cagagcggag ccccagtcgt gaatgaccag gacgacctct tccagaccca gggcttcgat
     4141 gaaggcatcc atgaagcgga cgtggtcgtc gaagaaataa cccaggtctg gtttgtcgga
     4201 tttgcccata ccgatcaggt ctggagcaat gcagcgatgg gtcggtgcaa catgcgggat
     4261 gatgttgcgc cacacgtagg aggaggtcgg gttaccgtgc aggaacagca caggggtgcc
     4321 atcgcgcgga ccaacatcga cgtagtgcat gcgctcgccc aggacttcca cataatgggg
     4381 gtcgaatgga aagccagtac cgatttctgc catggtggct ttgctagccc tatagtgagt
     4441 cgtattaata ctctagcctt aagagctgta attgaactgg gagtggacac ctgtggagag
     4501 aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca agagtcttct
     4561 ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa ccttgatact
     4621 tacctgccca gtgcctcacg accaacttct gcagcttaag ttcgagactg ttgtgtcaga
     4681 agcactgact gcgttagcaa tttaactgtg ataaactacc gcaataaagc ttctagtgat
     4741 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     4801 catttgcgtc aacggggcgg ggttattacg acattttgga aagtcccgtt gattttggtg
     4861 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     4921 ccgctatcca cgcccattgg tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     4981 taatacgtag atgtactgcc aagtaggaaa gtcccgtaag gtcatgtact gggcataatg
     5041 ccaggcgggc catttaccgt cattgacgtc aatagggggc ggacttggca tatgatacac
     5101 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     5161 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     5221 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     5281 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtgg cgcgccataa
     5341 tcctaggctc tcgtacgaag tcagctgata acgtcgacgt attgcggccg cggatcataa
     5401 t
//
