LOCUS Exported 9952 bp ds-DNA circular SYN 28-JUN-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS ED1L20-2 pSH-EFIRES-P-AtAFB2 new SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9952) AUTHORS . TITLE Direct Submission JOURNAL Exported Saturday, July 6, 2019 from SnapGene Viewer 3.1.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9952 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(306..894) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1065..1925) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(1926..2030) /gene="bla" /note="AmpR promoter" rep_origin 2057..2512 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" misc_feature 2642..3832 /note="AAVS1-LH" polyA_signal 3858..3906 /note="synthetic polyadenylation signal" misc_feature 3920..4011 /note="pause site" /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" promoter 4023..5185 /note="EF-1-alpha promoter" /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 4254..5176 /note="EF-1-alpha intron A" /note="intron upstream of the start codon of human EF-1-alpha" intron 5224..5356 /note="chimeric intron" /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" promoter 5401..5419 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 5446..7173 /note="AtAFB2" misc_feature 7209..7761 /note="IRES" /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 7761..8363 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /note="PuroR" /note="confers resistance to puromycin" /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" polyA_signal 8597..8678 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" misc_feature 8705..9946 /note="AAVS1-RH" ORIGIN 1 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta 61 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag 121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 481 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 721 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 961 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 1021 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 1081 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 1141 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 1201 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 1261 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 1321 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 1381 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 1441 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 1501 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 1561 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 1621 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 1681 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 1741 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 1801 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 1861 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 1921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 1981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 2041 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 2221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 2281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 2341 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 2401 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 2461 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg 2521 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 2581 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc 2641 gtcgacctac tctcttccgc attggagtcg ctttaactgg ccctggcttt ggcagcctgt 2701 gctgacccat gcagtcctcc ttaccatccc tccctcgact tcccctcttc cgatgttgag 2761 cccctccagc cggtcctgga ctttgtctcc ttccctgccc tgccctctcc tgaacctgag 2821 ccagctccca tagctcagtc tggtctatct gcctggccct ggccattgtc actttgcgct 2881 gccctcctct cgcccccgag tgcccttgct gtgccgccgg aactctgccc tctaacgctg 2941 ccgtctctct cctgagtccg gaccactttg agctctactg gcttctgcgc cgcctctggc 3001 ccactgtttc cccttcccag gcaggtcctg ctttctctga cctgcattct ctcccctggg 3061 cctgtgccgc tttctgtctg cagcttgtgg cctgggtcac ctctacggct ggcccagatc 3121 cttccctgcc gcctccttca ggttccgtct tcctccactc cctcttcccc ttgctctctg 3181 ctgtgttgct gcccaaggat gctctttccg gagcacttcc ttctcggcgc tgcaccacgt 3241 gatgtcctct gagcggatcc tccccgtgtc tgggtcctct ccgggcatct ctcctccctc 3301 acccaacccc atgccgtctt cactcgctgg gttccctttt ccttctcctt ctggggcctg 3361 tgccatctct cgtttcttag gatggccttc tccgacggat gtctcccttg cgtcccgcct 3421 ccccttcttg taggcctgca tcatcaccgt ttttctggac aaccccaaag taccccgtct 3481 ccctggcttt agccacctct ccatcctctt gctttctttg cctggacacc ccgttctcct 3541 gtggattcgg gtcacctctc actcctttca tttgggcagc tcccctaccc cccttacctc 3601 tctagtctgt gctagctctt ccagccccct gtcatggcat cttccagggg tccgagagct 3661 cagctagtct tcttcctcca acccgggccc ctatgtccac ttcaggacag catgtttgct 3721 gcctccaggg atcctgtgtc cccgagctgg gaccacctta tattcccagg gccggttaat 3781 gtggctctgg ttctgggtac ttttatctgt cccctccacc ccacagtggg gcgcatgcgt 3841 ggaattatca cctcgagaat aaaatatctt tattttcatt acatctgtgt gttggttttt 3901 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac 3961 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tactagtcgt 4021 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 4081 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa 4141 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 4201 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg 4261 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa 4321 ttacttccac ctggctccag tacgtgattc ttgatcccga gctggagcca ggggcgggcc 4381 ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg 4441 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct 4501 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt 4561 aaatgcgggc caggatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg 4621 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag 4681 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc 4741 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga 4801 aagatggccg cttcccggcc ctgctccagg gggctcaaaa tggaggacgc ggcgctcggg 4861 agagcgggcg ggtgagtcac ccacacaaag gaaaggggcc tttccgtcct cagccgtcgc 4921 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctggagctt 4981 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac 5041 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt 5101 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt 5161 ttttcttcca tttcaggtgt cgtgaacacg ggctgcagaa gttggtcgtg aggcactggg 5221 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg 5281 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt 5341 gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact 5401 taatacgact cactataggc tagcgctacc ggactcagat ctaccatgaa ctactttccc 5461 gatgaggtca ttgagcacgt ctttgatttc gtcacaagcc acaaggatag gaacgccatt 5521 agcctggtct gtaagagctg gtacaagatc gagcggtatt ccagacagaa ggtgttcatc 5581 ggcaactgct acgccatcaa tccagagagg ctgctgcgga gatttccctg tctgaagagc 5641 ctgaccctga agggcaagcc ccacttcgcc gactttaacc tggtgcctca cgagtgggga 5701 ggattcgtgc tgccatggat cgaggccctg gcacggtcca gagtgggcct ggaggagctg 5761 aggctgaagc gcatggtggt gacagacgag tctctggagc tgctgtctcg gagcttcgtg 5821 aacttcaaga gcctggtgct ggtgtcctgc gagggcttta ccacagatgg cctggcaagc 5881 atcgcagcaa actgtaggca cctgcgggac ctggacctgc aggagaatga gatcgacgat 5941 cacagaggcc agtggctgtc ctgcttcccc gatacctgta ccacactggt gacactgaac 6001 tttgcctgcc tggagggcga gacaaatctg gtggccctgg agaggctggt ggcacgctct 6061 cccaacctga agagcctgaa gctgaatagg gcagtgcctc tggacgcact ggcaagactg 6121 atggcatgcg caccacagat cgtggatctg ggcgtgggct cctacgagaa cgaccctgat 6181 tccgagtctt atctgaagct gatggccgtg atcaagaagt gtaccagcct gcgcagcctg 6241 tccggcttcc tggaggcagc acctcactgc ctgtccgcct ttcacccaat ctgtcacaac 6301 ctgacatccc tgaatctgtc ttacgccgcc gagatccacg gctcccacct gatcaagctg 6361 atccagcact gcaagaagct gcagaggctg tggatcctgg actccatcgg cgataagggc 6421 ctggaggtgg tggcctctac ctgtaaggag ctgcaggagc tgcgcgtgtt cccatctgac 6481 ctgctgggag gaggaaacac cgcagtgaca gaggagggcc tggtggcaat ctctgccgga 6541 tgccccaagc tgcacagcat cctgtatttt tgtcagcaga tgaccaacgc cgccctggtg 6601 acagtggcca agaactgccc caatttcatc aggtttcgcc tgtgcatcct ggagcccaat 6661 aagcctgacc acgtgacatc tcagcctctg gatgagggct tcggcgccat cgtgaaggcc 6721 tgcaagagcc tgaggcgcct gtctctgagc ggcctgctga ccgaccaggt gttcctgtac 6781 atcggcatgt atgccaacca gctggagatg ctgtctatcg cctttgccgg cgacacagat 6841 aagggcatgc tgtacgtgct gaatggctgt aagaagatga agaagctgga gatcagagac 6901 agcccttttg gcgataccgc cctgctggca gacgtgagca agtatgagac aatgcggagc 6961 ctgtggatga gctcctgcga ggtgaccctg agcggctgta agagactggc cgagaaggcc 7021 ccatggctga acgtggagat catcaacgag aatgacaaca ataggatgga ggagaatggc 7081 cacgagggcc gccagaaggt ggataagctg tatctgtata ggaccgtcgt cgggactcgg 7141 atggatgccc ccccctttgt ctggattctg taagcggccg ctctagccca attccgcccc 7201 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat 7261 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt 7321 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt 7381 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc 7441 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc 7501 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat 7561 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc 7621 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg 7681 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct 7741 ttgaaaaaca cgatgataat atgggcaccg agtacaagcc cacggtgcgc ctcgccaccc 7801 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca 7861 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct 7921 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg 7981 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc 8041 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc 8101 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg 8161 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc 8221 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc 8281 ggctcggctt caccgtcacc gccgacgtcg agtgcccgaa ggaccgcgcg acctggtgca 8341 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga 8401 gcgcacgacc ccatggctcc gaccgaagcc acccggggcg gccccgccga ccccgcaccc 8461 gcccccgagg cccaccgact ctagctagag gatcataatc agccatacca catttgtaga 8521 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa 8581 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag 8641 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccat 8701 gcatcactag ggacaggatt ggtgacagaa aagccccatc cttaggcctc ctccttccta 8761 gtctcctgat attgggtcta acccccacct cctgttaggc agattcctta tctggtgaca 8821 cacccccatt tcctggagcc atctctctcc ttgccagaac ctctaaggtt tgcttacgat 8881 ggagccagag aggatcctgg gagggagagc ttggcagggg gtgggaggga agggggggat 8941 gcgtgacctg cccggttctc agtggccacc ctgcgctacc ctctcccaga acctgagctg 9001 ctctgacgcg gctgtctggt gcgtttcact gatcctggtg ctgcagcttc cttacacttc 9061 ccaagaggag aagcagtttg gaaaaacaaa atcagaataa gttggtcctg agttctaact 9121 ttggctcttc acctttctag tccccaattt atattgttcc tccgtgcgtc agttttacct 9181 gtgagataag gccagtagcc agccccgtcc tggcagggct gtggtgagga ggggggtgtc 9241 cgtgtggaaa actccctttg tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg 9301 cctctactcc ctttctcttt ctccatcctt ctttccttaa agagtcccca gtgctatctg 9361 ggacatattc ctccgcccag agcagggtcc cgcttcccta aggccctgct ctgggcttct 9421 gggtttgagt ccttggcaag cccaggagag gcgctcaggc ttccctgtcc cccttcctcg 9481 tccaccatct catgcccctg gctctcctgc cccttcccta caggggttcc tggctctgct 9541 cttcagactg agccccgttc ccctgcatcc ccgttcccct gcatccccct tcccctgcat 9601 cccccagagg ccccaggcca cctacttggc ctggacccca cgagaggcca ccccagccct 9661 gtctaccagg ctgccttttg ggtggattct cctccaactg tggggtgact gcttggcaaa 9721 ctcactcttc ggggtatccc aggaggcctg gagcattggg gtgggctggg gttcagagag 9781 gagggattcc cttctcaggt tacgtggcca agaagcaggg gagctgggtt tgggtcaggt 9841 ctgggtgtgg ggtgaccagc ttatgctgtt tgcccaggac agcctagttt tagcgctgaa 9901 accctcagtc ctaggaaaac agggatggtt ggtcactgtc tctgggctcg ac //