LOCUS       sequence-471903-        6372 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express SELENOP-SmBiT-HaloTag(R) 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 6372)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6372)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SELENOP-SmBiT-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2307..2327
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2343..3227
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3315..3333)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3370..3491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3381..3400)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3435..3454
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3736..4093
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3736..3927
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3759..3779)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3944..4079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4108..4155
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4187..4981
                     /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(4241..4260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4851..4870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5045..5093
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5332..5920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5412..5431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6036..6319
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgtgg agaagcctgg
     1081 ggcttgccct ggctctctgt ctcctcccat cgggaggaac agagagccag gaccaaagct
     1141 ccttatgtaa gcaaccccca gcctggagca taagagatca agatccaatg ctaaactcca
     1201 atggttcagt gactgtggtt gctcttcttc aagccagctg atacctgtgc atactgcagg
     1261 catctaaatt agaagacctg cgagtaaaac tgaagaaaga aggatattct aatatttctt
     1321 atattgttgt taatcatcaa ggaatctctt ctcgattaaa atacacacat cttaagaata
     1381 aggtttcaga gcatattcct gtttatcaac aagaagaaaa ccaaacagat gtctggactc
     1441 ttttaaatgg aagcaaagat gacttcctca tatatgatag atgtggccgt cttgtatatc
     1501 atcttggttt gcctttttcc ttcctaactt tcccatatgt agaagaagcc attaagattg
     1561 cttactgtga aaagaaatgt ggaaactgct ctctcacgac tctcaaagat gaagactttt
     1621 gtaaacgtgt atctttggct actgtggata aaacagttga aactccatcg cctcattacc
     1681 atcatgagca tcatcacaat catggacatc agcaccttgg cagcagtgag ctttcagaga
     1741 atcagcaacc aggagcacca aatgctccta ctcatcctgc tcctccaggc cttcatcacc
     1801 accataagca caagggtcag cataggcagg gtcacccaga gaaccgagat atgccagcaa
     1861 gtgaagattt acaagattta caaaagaagc tctgtcgaaa gagatgtata aatcaattac
     1921 tctgtaaatt gcccacagat tcagagttgg ctcctaggag ctgatgctgc cattgtcgac
     1981 atctgatatt tgaaaaaaca gggtctgcaa tcacctgaca gtgtaaagaa aacctcccat
     2041 ctttatgtag ctgacaggga cttcgggcag aggagaacat aactgaatct tgtcagtgac
     2101 gtttgcctcc agctgcctga caaataagtc agcagcttat acccacagaa gccagtgcca
     2161 gttgacgctg aaagaatcag gcaaaaaagt gagaatgacc ttcaaacgtt tctcaaggca
     2221 gttcaggtgg tggcgggagc ggaggtggag gctcgagcgg tgtgaccggc taccggctgt
     2281 tcgaggagat tctcgagcca accactgagg atctgtactt tcagagcgat aacgatggat
     2341 ccgaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca
     2401 tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc
     2461 cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca
     2521 ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg
     2581 acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc
     2641 tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc
     2701 gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc
     2761 cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga
     2821 tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga
     2881 ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac
     2941 tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg
     3001 tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca
     3061 ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact
     3121 gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga
     3181 tcggcagcga gatcgcgcgc tggctgtcta ctctggagat ttccggttaa tagaattcta
     3241 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     3301 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag
     3361 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3421 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3481 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     3541 gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa
     3601 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3661 cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac
     3721 ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3781 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3841 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3901 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3961 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4021 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4081 aggcttttgc aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc
     4141 atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga
     4201 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4261 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4321 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4381 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4441 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4501 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4561 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4621 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4681 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4741 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4801 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4861 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4921 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4981 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     5041 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     5101 agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     5161 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacccggtat cagctcactc
     5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5761 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5881 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     5941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6001 atcaaaaagg atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg
     6061 atggcccttc gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg
     6121 ggtaatgtgg cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc
     6181 gcgattgcgg cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc
     6241 gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt
     6301 ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga
     6361 ggatcgagat ct
//
