LOCUS       sequence-479824-        5772 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express SmBiT-CYBB Fusion Protein in Mammalian Cells under a TK 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5772)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5772)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: SmBiT-CYBB (TK) Vector
FEATURES             Location/Qualifiers
     source          1..5772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..753
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(2715..2733)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2770..2891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2781..2800)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2835..2854
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3136..3493
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3136..3327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3159..3179)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3344..3479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3508..3555
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3587..4381
                     /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(3641..3660)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4251..4270
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4445..4493
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4732..5320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4812..4831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5436..5719
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 aaatgagtct tcggacctcg cgggggccgc ttaagcggtg gttagggttt gtctgacgcg
       61 gggggagggg gaaggaacga aacactctca ttcggaggcg gctcggggtt tggtcttggt
      121 ggccacgggc acgcagaaga gcgccgcgat cctcttaagc acccccccgc cctccgtgga
      181 ggcgggggtt tggtcggcgg gtggtaactg gcgggccgct gactcgggcg ggtcgcgcgc
      241 cccagagtgt gaccttttcg gtctgctcgc agacccccgg gcggcgccgc cgcggcggcg
      301 acgggctcgc tgggtcctag gctccatggg gaccgtatac gtggacaggc tctggagcat
      361 ccgcacgact gcggtgatat taccggagac cttctgcggg acgagccggg tcacgcggct
      421 gacgcggagc gtccgttggg cgacaaacac caggacgggg cacaggtaca ctatcttgtc
      481 acccggaggc gcgagggact gcaggagctt cagggagtgg cgcagctgct tcatccccgt
      541 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
      601 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaagtcga acacgcagat
      661 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
      721 gaacaccgag cgaccctgca gcgacccgct taaaagcttg gcaatccggt actgttggta
      781 aagccaccgc tagcgctcac catggtgacc ggctaccggc tgttcgagga gattctcggg
      841 agctccggtg gtggcgggag cggaggtgga ggctcgagcg gtgcgatcgc catggggaac
      901 tgggctgtga atgaggggct ctccattttt gtcattctgg tttggctggg gttgaacgtc
      961 ttcctctttg tctggtatta ccgggtttat gatattccac ctaagttctt ttacacaaga
     1021 aaacttcttg ggtcagcact ggcactggcc agggcccctg cagcctgcct gaatttcaac
     1081 tgcatgctga ttctcttgcc agtctgtcga aatctgctgt ccttcctcag gggttccagt
     1141 gcgtgctgct caacaagagt tcgaagacaa ctggacagga atctcacctt tcataaaatg
     1201 gtggcatgga tgattgcact tcactctgcg attcacacca ttgcacatct atttaatgtg
     1261 gaatggtgtg tgaatgcccg agtcaataat tctgatcctt attcagtagc actctctgaa
     1321 cttggagaca ggcaaaatga aagttatctc aattttgctc gaaagagaat aaagaaccct
     1381 gaaggaggcc tgtacctggc tgtgaccctg ttggcaggca tcactggagt tgtcatcacg
     1441 ctgtgcctca tattaattat cacttcctcc accaaaacca tccggaggtc ttactttgaa
     1501 gtcttttggt acacacatca tctctttgtg atcttcttca ttggccttgc catccatgga
     1561 gctgaacgaa ttgtacgtgg gcagaccgca gagagtttgg ctgtgcataa tataacagtt
     1621 tgtgaacaaa aaatctcaga atggggaaaa ataaaggaat gcccaatccc tcagtttgct
     1681 ggaaaccctc ctatgacttg gaaatggata gtgggtccca tgtttctgta tctctgtgag
     1741 aggttggtgc ggttttggcg atctcaacag aaggtggtca tcaccaaggt ggtcactcac
     1801 cctttcaaaa ccatcgagct acagatgaag aagaaggggt tcaaaatgga agtgggacaa
     1861 tacatttttg tcaagtgccc aaaggtgtcc aagctggagt ggcacccttt tacactgaca
     1921 tccgcccctg aggaagactt ctttagtatc catatccgca tcgttgggga ctggacagag
     1981 gggctgttca atgcttgtgg ctgtgataag caggagtttc aagatgcgtg gaaactacct
     2041 aagatagcgg ttgatgggcc ctttggcact gccagtgaag atgtgttcag ctatgaggtg
     2101 gtgatgttag tgggagcagg gattggggtc acacccttcg catccattct caagtcagtc
     2161 tggtacaaat attgcaataa cgccaccaat ctgaagctca aaaagatcta cttctactgg
     2221 ctgtgccggg acacacatgc ctttgagtgg tttgcagatc tgctgcaact gctggagagc
     2281 cagatgcagg aaaggaacaa tgccggcttc ctcagctaca acatctacct cactggctgg
     2341 gatgagtctc aggccaatca ctttgctgtg caccatgatg aggagaaaga tgtgatcaca
     2401 ggcctgaaac aaaagacttt gtatggacgg cccaactggg ataatgaatt caagacaatt
     2461 gcaagtcaac accctaatac cagaatagga gttttcctct gtggacctga agccttggct
     2521 gaaaccctga gtaaacaaag catctccaac tctgagtctg gccctcgggg agtgcatttc
     2581 attttcaaca aggaaaactt cgtttaaacg aattcgggct cggtacccgg ggatcctcta
     2641 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     2701 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag
     2761 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2821 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2881 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     2941 gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa
     3001 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3061 cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac
     3121 ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3241 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3301 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3361 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3421 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3481 aggcttttgc aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc
     3541 atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga
     3601 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3661 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3721 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3781 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3841 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3901 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3961 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4021 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4081 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4141 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4201 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4261 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4321 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4381 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     4441 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4501 agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     4561 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacccggtat cagctcactc
     4621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5161 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg
     5461 atggcccttc gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg
     5521 ggtaatgtgg cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc
     5581 gcgattgcgg cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc
     5641 gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt
     5701 ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga
     5761 ggatcgagat ct
//
