LOCUS pTwist_cDNA_BG 6525 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 6525) AUTHORS siwong TITLE Direct Submission JOURNAL Exported Oct 28, 2024 from SnapGene 7.2.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6525 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" intron 718..1193 /label=beta-globin intron /note="internally truncated intron from human beta-globin" misc_feature 1237^1238 /label=Your gene here misc_feature 1252..1840 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(1723..1734) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 1906..1954 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2156..2584 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2598..2927 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2778..2913 /label=SV40 ori /note="SV40 origin of replication" CDS 2994..3788 /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 3964..4085 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4134..4150) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4158..4174 /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(4182..4188,4189..4206,4207..4212)) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4227..4248 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4536..5124) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(join(5295..6086,6087..6155)) /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(6156..6260) /gene="bla" /label=AmpR promoter ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 661 ccagcctccc ctcgaagctt acatgtggta ccgagctcgg atcctgagaa cttcagggtg 721 agtctatggg acccttgatg ttttctttcc ccttcttttc tatggttaag ttcatgtcat 781 aggaagggga gaagtaacag ggtacacata ttgaccaaat cagggtaatt ttgcatttgt 841 aattttaaaa aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact 901 ttccctaatc tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca 961 ttctaaagaa taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa 1021 atatttctgc atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac 1081 aatccagcta ccattctgct tttattttat ggttgggata aggctggatt attctgagtc 1141 caagctaggc ccttttgcta atcatgttca tacctcttat cttcctccca cagctcctgg 1201 gcaacgtgct ggtctgtgtg ctggcccatc actttggggc cggccagatc taatcaacct 1261 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 1321 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 1381 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 1441 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 1501 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 1561 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 1621 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 1681 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 1741 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 1801 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa 1861 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 1921 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 1981 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 2041 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 2101 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg 2161 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 2221 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 2281 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct 2341 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 2401 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 2461 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 2521 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 2581 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 2641 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 2701 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 2761 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 2821 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 2881 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 2941 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3001 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3061 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3121 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3181 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3241 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3301 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3361 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3421 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3481 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3541 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3601 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3661 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3721 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 3781 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 3841 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 3901 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 3961 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4021 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4081 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 4141 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4201 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4261 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4321 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 4921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 4981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5221 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5401 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5581 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 5821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 5881 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 5941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6241 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 6301 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt 6361 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 6421 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta 6481 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt //