LOCUS       sequence-214904-        7262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector to test the effect of substituting 
            selenocysteine for Cys on luciferase enzyme activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7262)
  AUTHORS   Mehta A, Rebsch CM, Kinzy SA, Fletcher JE, Copeland PR
  TITLE     Efficiency of mammalian selenocysteine incorporation.
  JOURNAL   J Biol Chem. 2004 Sep 3;279(36):37852-9. doi: 
            10.1074/jbc.M404639200. Epub 2004 Jun 30.
  PUBMED    15229221
REFERENCE   2  (bases 1 to 7262)
  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: 
            "10.1074/jbc.M404639200"; journalName: "J Biol Chem"; date: 
            "2004-09-3- 3"; volume: "279"; issue: "36"; pages: "37852-9"
COMMENT     Sequence Label: LUC2 (L/Sec/deltaP)
FEATURES             Location/Qualifiers
     source          1..7262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             954..2606
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNMQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLI*GFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1020..1038)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2760..2801
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2811..2828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2851..2868)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2857..3081
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3127..3555
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3214..3233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3424..3445
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3564..3584)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3569..3898
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3749..3884
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3811..3830
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3965..4759
                     /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(4019..4038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4629..4648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4935..5056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4972..4991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5026..5045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5105..5121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5105..5121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5118..5140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5129..5145
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5153..5159,5160..5177,5178..5183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5198..5219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5336..5353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5507..6095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5587..5606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6266..7057,7058..7126))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6889..6908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7127..7231)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgtccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttatggaag
      961 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc tctagaggat ggaaccgctg
     1021 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
     1081 cagatgcaca tatcgaggtg aacatcacgt acgcggaata cttcgaaatg tccgttcggt
     1141 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
     1201 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
     1261 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgaac atttcgcagc
     1321 ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacatg caaaaaaaat
     1381 taccaataat ccagaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
     1441 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtac
     1501 cagagtcctt tgatcgtgac aaaacaattg cactgataat gaattcctct ggatctactg
     1561 ggttacctaa gggtgtggcc cttccgcata gaactgcctg cgtcagattc tcgcatgcca
     1621 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1681 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgagga tttcgagtcg
     1741 tcttaatgta tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc
     1801 aaagtgcgtt gctagtacca accctatttt cattcttcgc caaaagcact ctgattgaca
     1861 aatacgattt atctaattta cacgaaattg cttctggggg cgcacctctt tcgaaagaag
     1921 tcggggaagc ggttgcaaaa cgcttccatc ttccagggat acgacaagga tatgggctca
     1981 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     2041 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     2101 gcgttaatca gagaggcgaa ttatgtgtca gaggacctat gattatgtcc ggttatgtaa
     2161 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     2221 tagcttactg ggacgaagac gaacacttct tcatagttga ccgcttgaag tctttgatta
     2281 aatacaaagg atatcaggtg gcccccgctg aattggaatc gatattgtta caacacccca
     2341 acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     2401 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     2461 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     2521 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     2581 agaagggcgg aaagtccaaa ttgtaaaatg taactttaat taatggagcc ttccaccccg
     2641 gcactccggt ctgcctgaaa accagcccgc tggtggggca gtcccgagga cctggcgtgc
     2701 atccccgccg gaggaaggtc cagagcggcc gctcgagtct agagggcccg cggttcgaag
     2761 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc
     2821 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2881 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2941 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3001 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3061 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3121 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3181 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3241 tcgccacgtt cgccggcttt cctcgtcaag ctctaaatcg ggggctccct ttagggttcc
     3301 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3361 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3421 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3481 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3541 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3661 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3721 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3781 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3841 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3901 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3961 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4021 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4081 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4141 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4201 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4261 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4321 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4381 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4441 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4501 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4561 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4621 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4681 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4741 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     4801 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4861 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4921 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4981 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5041 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5101 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5161 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5221 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5281 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5341 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5401 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5941 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6301 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6361 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6421 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6481 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6541 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6601 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6661 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6721 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6781 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6841 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6901 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6961 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7021 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7081 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7141 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7201 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7261 tc
//
