LOCUS       sequence-203200-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged fragment (amino acids 1873-2025) of human Rev3
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  AUTHORS   Thanos Halazonetis
  TITLE     Expression plasmids for production of Rev7 and Champ1 proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5483)
  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            "Unpublished"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: pCDZ2FE_Rev3_1873-2025
FEATURES             Location/Qualifiers
     source          1..5483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        117..496
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        497..700
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     651..671
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     745..764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        745..763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             792..815
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             834..857
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1394..1411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1400..1624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1670..2098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1757..1776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1967..1988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2107..2127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2272..2406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2333..2352
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2468..2515
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2534..2908
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3038..3159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3075..3094)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3129..3148
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3208..3224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3208..3224)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3221..3243)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3232..3248
                     /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(3256..3262,3263..3280,3281..3286))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3301..3322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3439..3456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3610..4198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3690..3709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4369..5160,5161..5229))
                     /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     4992..5011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5230..5334)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5409..5428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
       61 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      121 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      181 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      241 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      541 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      601 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      661 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      721 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      781 gcgcgacgat ggactacaag gacgacgatg acaagctcac tggttctaat ggtgactaca
      841 aggatgacga tgacaagggt ggtaatggta atggtggtaa gcttggtcgt actgctaaca
      901 tcctgaaacc acttatgtcc cccccaagta gggaagaaat tatggcaact ttgttggatc
      961 atgacctgtc tgagactatt taccaggaac cattttgcag taatccttct gatgtaccag
     1021 aaaagcccag ggagattggt ggacggctcc tcatggtaga aactcgactt gcaaatgatc
     1081 tggctgagtt tgagggagac ttttccttgg aaggacttcg tctttggaaa acagcattct
     1141 cagcaatgac tcagaatcca aggccagggt caccccttcg cagtggccaa ggagttgtca
     1201 ataaagggtc aagtaatagc cctaagatgg ttgaagataa aaaaattgtg attatgcctt
     1261 gcaaatgtgc cccaagtcga caactggttc aagtgtggct tcaagccaaa gaagaatacg
     1321 aacgttccaa gaaactgcct aaaacctaag cggccgctcg agtctagagg gcccgtttaa
     1381 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     1621 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     1681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2041 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2101 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2161 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2221 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2281 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2341 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2401 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2461 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2521 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     2581 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     2641 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     2701 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     2761 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     2821 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     2881 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     2941 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3001 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     3061 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3121 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     3181 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3241 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3301 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3361 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3421 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3481 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3541 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4021 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4141 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4201 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4261 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4321 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4381 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4441 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4501 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4561 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4621 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4681 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4741 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4801 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4861 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4921 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4981 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5041 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5101 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5161 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5221 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5281 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5341 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     5401 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     5461 gtgttggagg tcgctgagta gtg
//
