pEZY3-SPT

LOCUS       pEZY3-SPT              7595 bp DNA     circular SYN 16-MAR-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7595)
  AUTHORS   Joe JMM Lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     ATTB1:GTTTGTACAAAAAAGCAGGC
FEATURES             Location/Qualifiers
     source          1..7595
                     /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"
     primer_bind     950..1012
                     /label=STP_BP_attB Insert.FOR
     protein_bind    950..974
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          977..1073
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      1143..1152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1149..2971
                     /codon_start=1
                     /label=SPT
                     /translation="MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEE
                     AFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKST
                     RRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESM
                     TSATFRDTNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIR
                     CLRMDDKSIRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFR
                     VYIPNKPSKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSC
                     RNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDG
                     PLTLVSYKPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTC
                     SRKTNRWPMALLYGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRK
                     RLEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKK
                     CKKVICREHNIDMCQSCF*GRNGRQNWGSRH"
     polyA_signal    2972..3093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2983..3002)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3037..3056
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3037..3056
                     /label=Primer 1
     primer_bind     complement(3053..3121)
                     /label=STP_BP_attB Insert.REV
     protein_bind    complement(3095..3119)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3148..3166)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3149..3166)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3186..3203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3192..3416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3462..3890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3549..3568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3759..3780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3899..3919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3904..4233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4084..4219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4146..4165
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4300..5094
                     /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(4354..4373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4964..4983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5268..5389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5268..5308
                     /label=STP_BP_attB Insert.REV
     primer_bind     complement(5305..5324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5305..5324)
                     /label=Primer 1
     primer_bind     5359..5378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5438..5454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5438..5454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5451..5473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5462..5478
                     /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(5486..5516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5531..5552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5669..5686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5840..6428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5920..5939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6599..7459)
                     /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     7222..7241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7460..7564)
                     /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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttgattaata caagtttgta
      961 caaaaaagca ggcttagtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg
     1021 tggtggtgca aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta
     1081 cggaagtgtt acttctgctc taaaagctgc ggaattgtac ccaattcgtt aaggccaaat
     1141 tggccaccat gggctctagc ctggacgacg agcacatcct gagcgccctg ctgcagagcg
     1201 acgacgaact ggtgggcgag gacagcgaca gcgaggtcag cgaccacgtg tccgaggacg
     1261 acgtgcagtc cgacaccgag gaagccttca tcgacgaggt gcacgaagtg cagcctacca
     1321 gcagcggctc cgagatcctg gacgagcaga acgtgatcga gcagcctggc agctccctgg
     1381 ccagcaacag aatcctgacc ctgccccaga gaaccatcag aggcaagaac aagcactgct
     1441 ggtccacctc caagagcacc aggcggagca gagtgtccgc cctgaacatc gtgcggagcc
     1501 agaggggccc caccagaatg tgcagaaaca tctacgaccc cctgctgtgc ttcaagctgt
     1561 tcttcaccga cgagatcatc agcgagatcg tgaagtggac caacgccgag atcagcctga
     1621 agaggcggga gagcatgacc agcgccacct tcagagacac caacgaggac gagatctacg
     1681 ccttcttcgg catcctggtg atgaccgccg tgagaaagga caaccacatg agcaccgacg
     1741 acctgttcga cagatccctg agcatggtgt acgtgtccgt gatgagcaga gacagattcg
     1801 acttcctgat cagatgcctg agaatggacg acaagagcat cagacccacc ctgcgggaga
     1861 acgacgtgtt cacccccgtg cggaagatct gggacctgtt catccaccag tgcatccaga
     1921 actacacccc tggcgcccac ctgaccatcg atgagcagct gctgggcttc agaggcagat
     1981 gccccttcag agtgtacatc cccaacaagc ccagcaagta cggcatcaag atcctgatga
     2041 tgtgcgacag cggcaccaag tacatgatca acggcatgcc ctacctgggc agaggcaccc
     2101 agacaaacgg cgtgcccctg ggcgagtact acgtgaaaga actgagcaag cctgtgcatg
     2161 gcagctgcag gaacatcacc tgcgacaact ggttcaccag catccccctg gccaagaacc
     2221 tgctgcagga accctacaag ctgaccatcg tgggcaccgt gcggagcaac aagcgggaga
     2281 tcccagaggt gctgaagaac agcagatcca gacctgtggg aacaagcatg ttctgcttcg
     2341 acggccccct gaccctggtg tcctacaagc ccaagcccgc caagatggtg tacctgctgt
     2401 ccagctgcga cgaggacgcc agcatcaacg agagcaccgg caagccccag atggtgatgt
     2461 actacaacca gaccaagggc ggcgtggaca ccctggacca gatgtgcagc gtgatgacct
     2521 gcagcagaaa gaccaacaga tggcccatgg ccctgctgta cggcatgatc aatatcgcct
     2581 gcatcaacag cttcatcatc tacagccaca acgtgtccag caagggcgag aaggtgcaga
     2641 gccggaagaa attcatgcgg aacctgtaca tgagcctgac ctccagcttc atgagaaaga
     2701 gactggaagc ccccaccctg aagagatacc tgcgggacaa catcagcaac atcctgccca
     2761 aggaagtgcc aggaacaagc gacgacagca ccgaggaacc cgtgatgaag aagaggacct
     2821 actgcaccta ctgtcccagc aagatcagaa gaaaggccaa cgccagctgc aagaaatgca
     2881 aaaaagtgat ctgccgggag cacaacatcg acatgtgcca gagctgtttc tgaggccgta
     2941 acggccgcca gaattgggga tccagacatg ataagataca ttgatgagtt tggacaaacc
     3001 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3061 tttgtaacca ttataagctg caataaacaa gttaacccag ctttcttgta caaagtggtc
     3121 ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     3181 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3241 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3301 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3361 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3421 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3721 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3781 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3841 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3901 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4081 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4141 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4201 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4261 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4321 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4381 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4441 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4501 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4561 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4621 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4681 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4741 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4801 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4861 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4921 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4981 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5041 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5101 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5161 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5221 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5281 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5341 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5401 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5461 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5521 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5581 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5641 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5701 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5761 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5821 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5881 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5941 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6001 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6061 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6121 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6181 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6241 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6301 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6361 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7561 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//