LOCUS       sequence-8815-ex        7694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3 converted Gateway Destination vector for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7694)
  AUTHORS   Guo F, Chiang MY, Wang Y, Zhang YZ
  TITLE     An in vitro recombination method to convert restriction- and 
            ligation-independent expression vectors.
  JOURNAL   Biotechnol J. 2008 Mar . 3(3):370-7.
  PUBMED    18064608
REFERENCE   2  (bases 1 to 7694)
  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: "Biotechnol 
            J. 2008 Mar . 3(3):370-7."
COMMENT     Sequence Label: pEZY3
FEATURES             Location/Qualifiers
     source          1..7694
                     /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"
     protein_bind    950..1074
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1503..1808)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1605..1624)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2153..2812)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2727..2746
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2813..2915)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3093..3217)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3246..3264)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3247..3264)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3284..3301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3290..3514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3560..3988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3647..3666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3857..3878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3997..4017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4002..4332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4183..4318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4245..4264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4399..5193
                     /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(4453..4472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5063..5082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5367..5488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5404..5423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5458..5477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5537..5553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5537..5553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5550..5572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5561..5577
                     /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(5585..5591,5592..5609,5610..5615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5630..5651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5768..5785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5939..6527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6019..6038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6698..7489,7490..7558))
                     /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     7321..7340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7559..7663)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttgattaata caagtttgta
      961 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     1021 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatgaatcaa
     1081 ctacttagat ggtattagtg acctgtagtc gaccgacagc cttccaaatg ttcttcgggt
     1141 gatgctgcca acttagtcga ccgacagcct tccaaatgtt cttctcaaac ggaatcgtcg
     1201 tatccagcct actcgctatt gtcctcaatg ccgtattaaa tcataaaaag aaataagaaa
     1261 aagaggtgcg agcctctttt ttgtgtgaca aaataaaaac atctacctat tcatatacgc
     1321 tagtgtcata gtcctgaaaa tcatctgcat caagaacaat ttcacaactc ttatactttt
     1381 ctcttacaag tcgttcggct tcatctggat tttcagcctc tatacttact aaacgtgata
     1441 aagtttctgt aatttctact gtatcgacct gcagactggc tgtgtataag ggagcctgac
     1501 atttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg
     1561 agatcagcca cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc
     1621 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct
     1681 gacagcagac gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata
     1741 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta
     1801 aactgcattt caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg
     1861 ggcgacctca gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg
     1921 ggcttcattc tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg
     1981 agcaactgat agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc
     2041 tttttgacat acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc
     2101 gcaaatacgc atactgttat ctggctttta gtaagccgga tccacgcggc gtttacgccc
     2161 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     2221 catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     2281 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     2341 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     2401 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     2461 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     2521 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     2581 tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     2641 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     2701 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
     2761 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     2821 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     2881 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
     2941 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
     3001 ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttagtcgact
     3061 acaggtcact aataccatct aagtagttga ttcatagtga ctggatatgt tgtgttttac
     3121 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     3181 tttacgtttc tcgttcagct ttcttgtaca aagtggtcct cgagcatgca tctagagggc
     3241 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     3301 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3361 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3421 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3481 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3541 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3601 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3661 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     3721 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3781 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3841 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3901 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     3961 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4021 gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4141 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4201 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4261 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4321 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4381 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4441 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4501 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4561 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4621 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4681 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4741 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4801 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4861 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4921 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4981 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5041 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5101 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5161 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5221 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5281 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5341 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5401 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5461 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5521 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5581 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5641 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5701 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5761 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5821 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6121 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6361 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6421 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7561 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7621 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7681 tgccacctga cgtc
//
