LOCUS       sequence-422373-        7683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The ORF of human Med25 cDNA amplified from the RT human liver RNA 
            was cloned into pGEMTE vector and then subclone at XhoI/XbaI sites 
            of the pcDNA3.2(+), the mammalian expression vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7683)
  AUTHORS   Zolfaghari R, Ross C
  TITLE     Zolfaghari and Ross plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7683)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1_hMed25
FEATURES             Location/Qualifiers
     source          1..7683
                     /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"
     regulatory      993..1002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3274..3291)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3280..3504
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3550..3978
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3637..3656)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3847..3868
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3987..4007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3992..4321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4172..4307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4234..4253
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4388..5182
                     /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(4442..4461)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5052..5071
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5356..5477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5393..5412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5447..5466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5526..5542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5526..5542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5539..5561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5550..5566
                     /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(5574..5580,5581..5598,5599..5604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5619..5640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5757..5774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5928..6516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6008..6027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6687..7478,7479..7547))
                     /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     7310..7329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7548..7652)
                     /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 atgcccagta 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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgctcgag ccgccaccat ggtccccggg tccgagggcc
     1021 cggcccgcgc cgggagcgtg gtggccgacg tggtgtttgt gattgagggt acggccaacc
     1081 tgggacccta cttcgagggg ctccgcaagc actacctgct cccggccatc gagtatttta
     1141 atggtggtcc tcctgctgag acggacttcg ggggagacta tggggggacc caatacagcc
     1201 tcgtggtgtt caacacagtg gactgcgctc ccgagtccta cgtacaatgt cacgctccca
     1261 ccagcagcgc ctatgagttt gtcacctggc tcgatggcat taagttcatg ggcgggggtg
     1321 gtgagagctg cagcctcatc gcggaaggac tcagcacagc cttgcagctg tttgatgact
     1381 tcaagaagat gcgcgagcag attggccaga cgcaccgggt ctgcctcctc atctgcaact
     1441 cacccccata cttgttgcct gctgttgaga gcaccacgta ctctggatgc acaactgaga
     1501 atcttgtgca gcagattggg gagcggggga tccacttctc cattgtgtct ccccggaagc
     1561 tgcctgcgct tcggcttctg tttgagaagg cagccccccc ggccttgctg gagccgctgc
     1621 agcctccgac agatgtgagc caggacccga ggcacatggt gctggttcgg ggactcgtgc
     1681 tgcctgttgg gggtggctca gccccaggcc ccctccagtc aaagcagcca gtccccctgc
     1741 ctcccgccgc accctcaggt gccactctct cagcagcccc ccagcagcct ctgccccccg
     1801 tccccccgca gtaccaggtt cccgggaacc tgagtgcagc tcaggtggcc gcgcagaatg
     1861 cagtggaggc tgccaagaac cagaaggctg ggctgggccc tcgcttctcg cccatcaccc
     1921 ctctccaaca agctgctccc ggagtgggtc cccccttcag ccaggcccca gctccccaac
     1981 tacccccagg accccctggc gcccccaagc caccacctgc ttcccagccc agtctggtct
     2041 ccactgtggc ccctggctcc ggcctggctc ccacggcaca gcccggggca ccgtccatgg
     2101 caggcactgt ggccccagga ggggtgagcg gcccttcccc agcccagctg ggagccccag
     2161 ccctcggtgg gcagcagtca gtctccaata agcttctggc ctggagcggg gtcctggagt
     2221 ggcaagagaa acccaaacct gcctcagtgg atgccaacac caagctgacg cggtcactgc
     2281 cctgccaggt ctacgtgaat catggcgaga acctgaagac ggagcagtgg ccccagaagc
     2341 tgatcatgca gctcatcccc cagcagctgc tgaccaccct gggccctttg ttccggaact
     2401 caaggatggt ccagttccat ttcaccaaca aggacctgga gtctctcaaa ggcctctacc
     2461 gcatcatggg caacggcttc gcgggctgcg tgcacttccc ccacacggcg ccctgtgagg
     2521 tgcgcgtgct catgctcctg tactcgtcca agaagaagat cttcatgggc ctcatcccct
     2581 acgaccagag cggcttcgtc aacggcatcc ggcaggtcat caccaaccac aagcaggtcc
     2641 agcagcagaa gctggagcag cagcagcgag gaatgggggg acagcaggca cccccagggc
     2701 tggggcccat tctggaggac caagccaggc cctcacagaa tctgctccag ctccgcccac
     2761 cgcagcccca gcctcagggt accgtagggg cctctggggc cacggggcag ccccagcccc
     2821 aaggtactgc ccagcccccg ccaggtgccc ctcaaggccc tcctggagca gcttctggcc
     2881 caccccctcc tggacccatc cttcggcccc agaaccctgg ggccaaccct cagctgcgaa
     2941 gcctcctcct caacccacca ccgccgcaga ctggggtgcc cccaccccag gcctccctcc
     3001 accacctcca gccaccaggg gctcctgcgc tgctgcctcc gccgcaccag ggcctggggc
     3061 agccccagtt ggggccccca ctcctgcatc caccacctgc ccagtcctgg cccgcacaac
     3121 ttccccctcg ggctccactg ccaggtcaga tgctgctgag cgggggtccc cggggcccgg
     3181 tcccccagcc gggcctgcag cccagcgtca tggaggacga catcctcatg gatctcatct
     3241 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3301 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3361 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3421 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3481 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3541 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3601 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3661 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3721 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3781 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3841 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3901 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3961 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4021 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4081 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4141 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4201 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4261 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4321 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4381 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4441 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4501 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4561 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4621 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4681 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4741 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4801 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4861 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4921 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4981 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5041 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5101 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5161 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5221 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5281 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5341 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5401 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5461 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5521 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5581 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5641 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5701 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6361 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6961 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7621 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7681 gtc
//
