LOCUS       sequence-439441-        7150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged Psmc2-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7150)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7150)
  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"
COMMENT     Sequence Label: PSMC2_GFP_Cterm
FEATURES             Location/Qualifiers
     source          1..7150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    111..335
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     324..341
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(385..1098,1099..1101,1102..1104))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     779..798
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1038..1059
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1117..1137)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2550..2574)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2584..2602
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2605..2623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(2625..2828)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2654..2674)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2829..3208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3380..3399
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3474..3578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3579..3647,3648..4439)
                     /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     complement(3797..3816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4610..5198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5099..5118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5352..5369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5486..5507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5522..5527,5528..5545,5546..5552)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5560..5576
                     /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)."
     primer_bind     5565..5587
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5584..5600
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5584..5600
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5649..5770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5660..5679)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5714..5733
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5900..6922)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(5900..6922)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(6930..6977)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     7109..7128
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
       61 cgcgtgggga taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc
      121 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
      181 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
      241 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
      301 gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc gggtttaaac
      361 gggccctcta gactcgagtc actattactt gtacagctcg tccatgccga gagtgatccc
      421 ggcggcggtc acgaactcct taaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
      481 cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
      541 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
      601 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
      661 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
      721 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
      781 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
      841 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
      901 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
      961 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     1021 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     1081 cagctcctcg cccttgctca ccatggtggt accttcacct tgaaaatata aattttcgcg
     1141 gccgcggcag ttgtatgtca tgtaacgagg agtagcactg aatttggcat aagacttaat
     1201 gaccttattt acagcttcca agaaatcctt ctcggtagca atttttcgcc gtgctctgat
     1261 ggcaaacata ccagcctctg tgcagacgct tctaatctca gcaccagtgc tatttggaca
     1321 cagtcgtgct aacagttcaa atctgatatc tctttcaaca ctcattgaac gagcgtgaat
     1381 cttaaatatg tgggtccgac cctctagatc gggcaagcta aattcaattt ttctatccaa
     1441 tctccctggc ctcatcagtg ctggatccaa agtatcaggt ctgttagtgg ccatcagcac
     1501 tttaatattg cctcgaggat caaaaccatc aagctgattg atcagttcca acattgttct
     1561 ctgcacttca ttgtcacctc cagcaccatc atcaaaacga gcccctccaa tagcatcaat
     1621 ttcatcaaag aagataaggc aggctttttt tgttctggcc atttcaaaga gttcacgaac
     1681 cattcgagcc ccctcaccga cgtatttctg tacaagctca gatccaataa ctcgaatgaa
     1741 gcacgcatca gtccgattag caactgcccg cgcacagagt gtcttgcctg taccgggtgg
     1801 accaaagagc agcacgccct tgggaggctc aatgccaagg ttcacaaacc tctctggatg
     1861 aagtaatggg gtttcaacta cttctcgcag tttctcaatc tgttccttac agccaccaac
     1921 atcactgtat gtgacatcag gtttctcttc cacctgcatc atggtaactg ttgggtcaat
     1981 cttaggaggc aatggaatgt gaatttgata tttatttcta tccacgccca ctctcatccc
     2041 ttcttcaatg tcagtaggtg ccacctgatc actaaggtcc accacaaact tggcaaactg
     2101 ctttacgttg ataatgtatt ttgggtcctc cgaatcagca ttgattatct ttgtacacct
     2161 ggcaacctgt aaaggctgtt cactctggag tgtctgctta tctgcagcca aatcccagag
     2221 tgctggtggg gccaggccag tgtcagattc tttaatacca gtgagctcat taattttctt
     2281 gagaagttgc tgaatgtcat cttcaacttg cttgatctgc ctagagtaag tgctctgacc
     2341 ataagttttc aacaaggcaa tatccccctc atccagagct cggatgggct tgtcgtcctt
     2401 ctcatcctct ttggtcttcc gctgatcggc accgaggtaa tccggcatgc aggcctccta
     2461 agcttaagtt taaacgctag agtccggagg ctggatcggt cccggtgtct tctatggagg
     2521 tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg agctcgtcga
     2581 cgatctctat cactgatagg gagatctcta tcactgatag ggagagctct gcttatatag
     2641 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     2701 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     2761 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     2821 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     2881 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     2941 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     3001 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     3061 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     3121 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     3181 tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc
     3241 agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt
     3301 gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc
     3361 agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga
     3421 tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3481 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3541 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3601 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3661 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3721 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3781 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3841 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3901 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3961 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4021 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4081 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4141 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4201 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4261 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4321 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4381 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4441 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4501 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4561 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4621 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4681 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4741 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4801 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4861 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4921 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4981 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5041 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5101 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5161 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5221 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5281 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5341 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5401 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5461 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5521 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5581 acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     5641 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5701 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5761 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     5821 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     5881 cgaaatctcg tagtacgtgc tattcctttg ccctcggacg agtgctgggg cgtcggtttc
     5941 cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga
     6001 tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg
     6061 cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa
     6121 tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga
     6181 agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga
     6241 cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc
     6301 cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg
     6361 ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg
     6421 tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat
     6481 cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct
     6541 ggctaagatc ggccgcagcg atcgcatcca tggcctccgc gaccggctgc agaacagcgg
     6601 gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat
     6661 aggtcaggct ctcgctgaat tccccaatgt caagcacttc cggaatcggg agcgcggccg
     6721 atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc
     6781 gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg
     6841 agagctgcat caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg
     6901 tcgcggtgag ttcaggcttt ttggccaagg aagttcctat actttctaga gaataggaac
     6961 ttcggaatag gaacttctag gtacgtgaac catcacccta atcaagtttt ttggggtcga
     7021 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     7081 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     7141 cgctggcaag
//
