LOCUS Exported 7956 bp ds-DNA circular SYN 19-DEC-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7956) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Apr 21, 2022 from SnapGene 5.1.7 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7956 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 663..1679 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" protein_bind 1754..1787 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 1837..2631 /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" polyA_signal 2811..2945 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 2959..2992 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 3060..3069 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 3066..4450 /label=mRARa misc_feature 4458..4839 /label=VP16 polyA_site 5011..5066 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(5427..5443) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5451..5467 /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(5475..5505) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5520..5541 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." polyA_signal 5780..5914 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(6153..6741) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6912..7772) /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" promoter complement(7773..7877) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca 421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag 481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc 541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt 601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg 661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc 721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc 781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg 841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt 901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc 961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg 1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg 1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc 1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc 1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg 1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg 1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg 1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca 1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg 1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc 1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg 1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc 1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgattaatt 1741 cgagcgaacg cgtataactt cgtatagcat acattatacg aagttatctc gagtcggatt 1801 tgatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 1861 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 1921 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 1981 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 2041 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 2101 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 2161 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 2221 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 2281 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 2341 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 2401 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 2461 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 2521 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 2581 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 2641 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 2701 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 2761 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccatcgat aacttgttta 2821 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 2881 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 2941 ggatcaatcc gaacgcgtat aacttcgtat agcatacatt atacgaagtt atctcgagtc 3001 gctcggtacg atttaaattg aattctgcag tcgacggtac cgcgggcccg ggatccaccg 3061 ccaccatggc cagcaatagc agttcctgcc caacacctgg gggcgggcac ctcaatgggt 3121 acccagtacc cccctacgcc ttcttctttc cccctatgct gggtggactc tccccacccg 3181 gcgctctcac cagcctccag caccagcttc cagtcagtgg ttacagcaca ccgtccccag 3241 ccaccatcga gacccagagc agcagttccg aagagatagt acccagccct ccctcaccac 3301 cgcccctgcc ccgcatctac aagccttgct ttgtttgtca agacaaatca tccggctacc 3361 actatggggt cagtgcctgt gagggctgta agggcttctt ccgacgaagc atccagaaga 3421 acatggtgta tacgtgtcac cgggacaaga actgcatcat caacaaggtg acccggaacc 3481 gctgccagta ctgccggctg cagaaatgtt tcgacgtggg catgtcccag gagtcggtgc 3541 gaaacgatcg aaacaaaaag aagaaagagg cacccaagcc cgagtgctca gagagctaca 3601 cgctgacgcc tgaggtgggc gagctcattg agaaggttcg caaagcgcac caggagacct 3661 tcccggccct ctgccagctg ggcaagtaca ctacgaacaa cagctcagaa caacgagtct 3721 ccctggacat tgacctctgg gacaagttca gtgaactctc caccaagtgc atcattaaga 3781 ctgtggagtt cgccaagcag cttcccggct tcaccaccct caccatcgcc gaccagatca 3841 ccctcctcaa ggctgcctgc ctggatatcc tgattctgcg aatctgcacg cggtacacgc 3901 ctgagcaaga cacaatgacc ttctcagatg gactgaccct gaaccggact cagatgcaca 3961 acgctggctt tggccccctc accgacttgg tctttgcctt cgccaaccag ctgctgcccc 4021 tggagatgga cgatgctgag actggactgc tcagtgccat ctgcctcatc tgtggagacc 4081 gacaggacct ggagcagcca gacaaggtgg acatgctgca agagccgctg ctggaagcac 4141 tgaaagtcta cgtccggaaa cggaggccca gccgacccca catgttcccc aagatgctga 4201 tgaagatcac agaccttcgg agcatcagcg ccaagggagc tgaacgggtg atcacattga 4261 agatggagat cccaggctcc atgccaccgc tgatccagga aatgctggag aactctgagg 4321 gcttggacac tctaagcgga cagtcggggg gcggaacacg agatgggggt ggcctggccc 4381 cccctccggg tagctgtagc cccagcctca gtcccagctc ccacagaagc agcccagcca 4441 cccaatcccc aggaccggtc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg 4501 gcctgctcga tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc 4561 tgtcctttct ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca 4621 gcctggggga cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc 4681 tagacgattt cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc 4741 cccacgactc cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt 4801 ttaccgatgc ccttggaatt gacgagtacg gtgggtaggc ggccgctctc ctcacgtgca 4861 ggctgtgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg 4921 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca 4981 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag 5041 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa 5101 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca 5161 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt 5221 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt 5281 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc 5341 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc 5401 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 5461 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 5521 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 5581 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc 5641 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 5701 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 5761 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa 5821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 5881 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc 5941 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 6001 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 6061 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 6121 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 6181 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 6241 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6301 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6361 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6421 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 6481 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 6541 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 6601 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6661 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 6721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 6781 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 6841 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 6901 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 6961 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 7021 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 7081 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 7141 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 7201 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 7261 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 7321 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 7381 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 7441 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 7501 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 7561 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 7621 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 7681 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 7741 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 7801 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 7861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt 7921 gactagttat taatagtaat caattacggg gtcatt //