LOCUS       sequence-214894-        7931 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 7931)
  AUTHORS   Shetty SP, Shah R, Copeland PR
  TITLE     Regulation of selenocysteine incorporation into the selenium 
            transport protein, selenoprotein P.
  JOURNAL   J Biol Chem. 2014 Sep 5;289(36):25317-26. doi: 
            10.1074/jbc.M114.590430. Epub 2014 Jul 25.
  PUBMED    25063811
REFERENCE   2  (bases 1 to 7931)
  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.M114.590430"; journalName: "J Biol Chem"; date: 
            "2014-09-5- 5"; volume: "289"; issue: "36"; pages: "25317-26"
COMMENT     Sequence Label: LUC56 (Luc/UGA/SP)
FEATURES             Location/Qualifiers
     source          1..7931
                     /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             3429..3470
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3480..3497
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3520..3537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3526..3750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3796..4224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3883..3902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4093..4114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4233..4253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4238..4567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4418..4553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4480..4499
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4634..5428
                     /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(4688..4707)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5298..5317
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5604..5725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5641..5660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5695..5714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5774..5790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5774..5790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5787..5809)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5798..5814
                     /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(5822..5828,5829..5846,5847..5852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5867..5888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6005..6022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6176..6764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6256..6275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6935..7726,7727..7795))
                     /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     7558..7577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7796..7900)
                     /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 taaatattta aaataaagcg
     2641 tactctcccc aactccatct aaagacacaa tttcatttct agaatgtttc caatccatct
     2701 aattaattag tgaagtaaaa gtagttgaaa ttggatttgt gcaaacatgg agaaatctac
     2761 cacattggct tctaaaattt aaaattttta tgccacaaac catttcatcc aaatcagatt
     2821 tgtaccgtgg ggcaactgaa aagtgattgc ggccattggt taatatgtct tcctttttct
     2881 ttctccagtg ttctagttac attgatgaga acagaaacat aaactatgac ctaggggttt
     2941 ctgttggata gctcgtaatt aagaacggag aaagaacaac aaagacatat tttccagttt
     3001 tttttcttta cttaaaactt tcaaaacaat agaaactttg tctttctaat cttatacttt
     3061 aaaccgatta aatctttaac agactacatt ttaaatatct acttatcttt tttatctcta
     3121 agactcctag tttgagtttc actacatata tctgtgaatc ttgttttttt catctaatgc
     3181 tgtatcagtc ttctgagttg tgagtgactg tcttgaaaga tgtaatggaa gaaaagtatg
     3241 gtgttaatct gcatagtgct taagacagta tttccataat caatgacggt ttaatagaga
     3301 aactgagtcc tatgaacctg aactccttta tggctaatac aattaagcaa gaatggagaa
     3361 tagaattgat tggctacagt acagattatc aaagcggccg ctcgagtcta gagggcccgc
     3421 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc
     3481 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3721 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3781 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3841 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3901 cttcctttct cgccacgttc gccggctttc ctcgtcaagc tctaaatcgg gggctccctt
     3961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4141 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4201 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4321 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4381 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4441 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4501 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4561 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4621 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4681 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4741 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4801 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4861 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4921 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4981 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5041 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5101 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5161 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5221 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5281 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5341 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5401 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     5461 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5521 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5581 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5641 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5701 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5761 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5821 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5881 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5941 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6001 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6061 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6121 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6361 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6601 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6661 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7921 cacctgacgt c
//
