LOCUS pTwist_cDNA_EF1a 6508 bp DNA circular SYN 22-OCT-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6508) AUTHORS siwong TITLE Direct Submission JOURNAL Exported Oct 28, 2024 from SnapGene 7.2.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6508 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..1178 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 232..1170 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" misc_feature 1197^1198 /label=Your gene here misc_feature 1235..1823 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(1706..1717) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 1889..1937 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2139..2567 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2581..2910 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2761..2896 /label=SV40 ori /note="SV40 origin of replication" CDS 2977..3771 /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 3947..4068 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4117..4133) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4141..4157 /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(4165..4171,4172..4189,4190..4195)) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4210..4231 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4519..5107) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(join(5278..6069,6070..6138)) /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(6139..6243) /gene="bla" /label=AmpR promoter ORIGIN 1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc 241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt 301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 721 ggtctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgga gacccaagct ggctagctct 1201 agagggcccg tttaaacccg ctgatcagcc tcgaaatcaa cctctggatt acaaaatttg 1261 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 1321 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 1381 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 1441 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 1501 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 1561 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 1621 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 1681 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 1741 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 1801 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag 1861 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg 1921 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac 1981 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc 2041 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat 2101 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 2161 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 2221 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 2281 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 2341 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 2401 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 2461 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc 2521 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 2581 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 2641 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 2701 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 2761 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 2821 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 2881 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 2941 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3001 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3061 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3121 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 3181 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3241 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3301 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3361 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 3421 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 3481 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 3541 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 3601 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 3661 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 3721 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 3781 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 3841 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 3901 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 3961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 4021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 4081 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 4141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 4201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 4261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4921 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 4981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 5041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 5221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 5881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 6241 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc 6301 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc 6361 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg 6421 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt 6481 cgcgatgtac gggccagata tacgcgtt //