LOCUS       sequence-203204-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged fragment (amino acids 1873-2025) of human Rev3
            protein with mutations abolishing Rev7 binding.
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  JOURNAL   Unpublished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            "Unpublished"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_4PA
FEATURES             Location/Qualifiers
     source          1..5483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        494..697
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     648..668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     742..761
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        742..760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             789..812
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             831..854
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1391..1408)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1397..1621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1667..2095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1754..1773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1964..1985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2104..2124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2269..2403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2330..2349
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2465..2512
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2531..2905
                     /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    3035..3156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3072..3091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3126..3145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3205..3221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3205..3221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3218..3240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3229..3245
                     /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(3253..3259,3260..3277,3278..3283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3298..3319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3436..3453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3607..4195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3687..3706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4366..5157,5158..5226))
                     /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     4989..5008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5227..5331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5406..5425)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
       61 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      121 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      181 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      241 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      301 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      361 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      421 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      481 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      541 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      601 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      661 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      721 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg
      781 cgacgatgga ctacaaggac gacgatgaca agctcactgg ttctaatggt gactacaagg
      841 atgacgatga caagggtggt aatggtaatg gtggtaagct tggtcgtact gctaacatcc
      901 tgaaagcact tatgtccccc gcaagtaggg aagaaattat ggcaactttg ttggatcatg
      961 acctgtctga gactatttac caggaaccat tttgcagtaa tccttctgat gtaccagaaa
     1021 agcccaggga gattggtgga cggctcctca tggtagaaac tcgacttgca aatgatctgg
     1081 ctgagtttga gggagacttt tccttggaag gacttcgtct ttggaaaaca gcattctcag
     1141 caatgactca gaatccaagg ccagggtcac cccttcgcag tggccaagga gttgtcaata
     1201 aagggtcaag taatagccct aagatggttg aagataaaaa aattgtgatt atggcttgca
     1261 aatgtgccgc aagtcgacaa ctggttcaag tgtggcttca agccaaagaa gaatacgaac
     1321 gttccaagaa actgcctaaa acctaagcgg ccgctcgagt ctagagggcc cgtttaaacc
     1381 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1441 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1501 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1561 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1621 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1681 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1741 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1801 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1861 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1921 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1981 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2041 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2101 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2161 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2281 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2341 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2401 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     2461 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     2521 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     2581 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     2641 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     2701 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     2761 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     2821 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     2881 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     2941 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3001 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3061 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3121 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3181 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4021 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5341 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     5401 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     5461 ttggaggtcg ctgagtagtg cgc
//
