LOCUS Exported 6168 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.1-SC35-cMyc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6168) AUTHORS Shi J, Hu Z, Pabon K, Scotto KW TITLE Caffeine regulates alternative splicing in a subset of cancer-associated genes: a role for SC35. JOURNAL Mol Cell Biol. 2008 Jan;28(2):883-95. Epub 2007 Nov 19. PUBMED 18025108 REFERENCE 2 (bases 1 to 6168) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6168 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1653..1682 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1698..1715 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1759..1776) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1765..1989 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2035..2463 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2122..2141) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2332..2353 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2472..2492) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2477..2806 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2657..2792 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2719..2738 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2873..3667 /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" primer_bind complement(2927..2946) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3537..3556 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3841..3962 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3878..3897) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3932..3951 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4011..4027) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4011..4027) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4024..4046) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4035..4051 /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(4059..4089) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4104..4125 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4242..4259) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4413..5001) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4493..4512) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5172..6032) /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 5795..5814 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6033..6137) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattca 961 tgagctacgg ccgcccccct cccgatgtgg agggtatgac ctccctcaag gtggacaacc 1021 tgacctaccg cacctcgccc gacacgctga ggcgcgtctt cgagaagtac gggcgcgtcg 1081 gcgacgtgta catcccgcgg gaccgctaca ccaaggagtc ccgcggcttc gccttcgttc 1141 gctttcacga caagcgcgac gctgaggacg ctatggatgc catggacggg gccgtgctgg 1201 acggccgcga gctgcgggtg caaatggcgc gctacggccg ccccccggac tcacaccaca 1261 gccgccgggg accgccaccc cgcaggtacg ggggcggtgg ctacggacgc cggagccgca 1321 gccctaggcg gcgtcgccgc agccgatccc ggagtcggag ccgttccagg tctcgcagcc 1381 gatctcgcta cagccgctcg aagtctcggt cccgcactcg ttctcgatct cggtcgacct 1441 ccaagtccag atccgcacga aggtccaagt ccaagtcctc gtcggtctcc agatctcgtt 1501 cgcggtccag gtcccggtct cggtccagga gtcctccccc agtgtccaag agggaatcca 1561 aatccaggtc gcgatcgaag agtcccccca agtctcctga agaggaagga gcggtgtcct 1621 ctttggatcc gagctcggta ccaagctttc tagaacaaaa actcatctca gaagaggatc 1681 tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttaaacggtc tccagcttaa 1741 gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 1801 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 1861 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 1921 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 1981 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 2041 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 2101 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 2161 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 2221 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 2281 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 2341 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 2401 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 2461 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 2521 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 2581 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 2641 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 2701 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 2761 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 2821 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 2881 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 2941 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 3001 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 3061 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 3121 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 3181 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 3241 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 3301 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 3361 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 3421 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 3481 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 3541 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 3601 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 3661 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 3721 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 3781 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 3841 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 3901 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 3961 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 4021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 4081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 4141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 4261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 4501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 4561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 4621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 4681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 4741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 4801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 4861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 4921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 4981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 5101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 5161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 5221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 5281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 5341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 5401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 5461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 5521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 5581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 5641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 5701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 5761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 5821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 5881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 5941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 6001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 6061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 6121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc //