LOCUS Exported 6357 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS PGKneolox2DTA.2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6357) TITLE Soriano vectors REFERENCE 2 (bases 1 to 6357) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6357 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 4..459 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(91..110) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 301..322 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 585..607 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 599..616 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 626..645 /label=T7 /note="T7 promoter, forward primer" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 727..743 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 727..743 /label=pBluescriptSK /note="For pBluescript vector" protein_bind complement(757..790) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 820..1319 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 1333..2136 /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1396..1415) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2006..2025 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2174..2398 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind complement(2457..2490) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 2569..3068 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 3095..3667 /codon_start=1 /gene="Corynebacterium diphtheriae tox" /product="diphtheria toxin fragment A" /label=DTA /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI NFETRGKRGQDAMYEYMAQACAGNRVRR" intron 3744..3809 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" polyA_signal 3883..4107 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4169..4189) /label=T3 /note="T3 promoter, forward primer" promoter complement(4169..4187) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4208..4224) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4208..4224) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4221..4243) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4232..4248 /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(4256..4286) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4301..4322 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4439..4456) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4610..5198) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4690..4709) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5369..6229) /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 5992..6011 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6230..6334) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg 481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca 661 ccgcggtggc ggccgcctag gagatctacg cgtcaattga tgcatcccgg gtttaaacta 721 ggccgctcta gaactagtgg atccggaacc cttaatataa cttcgtataa tgtatgctat 781 acgaagttat taggtccctc gacctgcagg aattctaccg ggtaggggag gcgcttttcc 841 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg 901 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg 961 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc 1021 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga 1081 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc 1141 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca 1201 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca 1261 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga 1321 cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc 1381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 1441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 1501 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 1561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 1621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 1681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 1741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 1801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 1861 ctcaaggcgc gcatgcccga cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg 1921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 1981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 2041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 2101 atcgccttct atcgccttct tgacgagttc ttctgagggg atcaattctc tagagctcgc 2161 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 2221 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 2281 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 2341 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 2401 tctgaggcgg aaagaaccag ctggggctcg aatcaagctg atccggaacc cttaatataa 2461 cttcgtataa tgtatgctat acgaagttat taggtccctc gacctgcagc ccaagctagc 2521 ttatcgatac cgtcgacggt atcgataagc ttgatatcga attctaccgg gtaggggagg 2581 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct 2641 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc 2701 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc 2761 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc 2821 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc 2881 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg 2941 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg 3001 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg 3061 gcctttcgac ctgcaggtcc tcgccatgga tcctgatgat gttgttgatt cttctaaatc 3121 ttttgtgatg gaaaactttt cttcgtacca cgggactaaa cctggttatg tagattccat 3181 tcaaaaaggt atacaaaagc caaaatctgg tacacaagga aattatgacg atgattggaa 3241 agggttttat agtaccgaca ataaatacga cgctgcggga tactctgtag ataatgaaaa 3301 cccgctctct ggaaaagctg gaggcgtggt caaagtgacg tatccaggac tgacgaaggt 3361 tctcgcacta aaagtggata atgccgaaac tattaagaaa gagttaggtt taagtctcac 3421 tgaaccgttg atggagcaag tcggaacgga agagtttatc aaaaggttcg gtgatggtgc 3481 ttcgcgtgta gtgctcagcc ttcccttcgc tgaggggagt tctagcgttg aatatattaa 3541 taactgggaa caggcgaaag cgttaagcgt agaacttgag attaattttg aaacccgtgg 3601 aaaacgtggc caagatgcga tgtatgagta tatggctcaa gcctgtgcag gaaatcgtgt 3661 caggcgatct ctttgtgaag gaaccttact tctgtggtgt gacataattg gacaaactac 3721 ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac 3781 tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc 3841 agtggtggaa tgcagatcct agagctcgct gatcagcctc gactgtgcct tctagttgcc 3901 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca 3961 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta 4021 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc 4081 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga 4141 gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt 4201 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 4261 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 4321 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 4381 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 4441 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4501 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4561 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4621 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4681 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4801 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 5041 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 5101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 5161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5341 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5401 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5461 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5521 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5581 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5641 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 5701 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5761 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 5821 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 5881 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 5941 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6001 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 6061 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 6121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac //