LOCUS Exported 4794 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION TOPO construct expressing UBC for generation of GMAP-compatible promoter. ACCESSION . VERSION . KEYWORDS TOPO UBC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4794) AUTHORS Akama-Garren EH, Joshi NS, Tammela T, Chang GP, Wagner BL, Lee DY, Rideout Iii WM, Papagiannakopoulos T, Xue W, Jacks T TITLE A Modular Assembly Platform for Rapid Generation of DNA Constructs. JOURNAL Sci Rep. 2016 Feb 18;6:16836. doi: 10.1038/srep16836. PUBMED 26887506 REFERENCE 2 (bases 1 to 4794) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4794 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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)." primer_bind 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 239..257 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 239..256 /label=SP6 /note="SP6 promoter, forward primer" promoter 330..1541 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 1378..1397 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" primer_bind complement(1681..1700) /label=T7 /note="T7 promoter, forward primer" promoter complement(1682..1700) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1707..1724) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1707..1723) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1716..1738) /label=M13/pUC Forward /note="In lacZ gene" CDS 1861..2163 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 2042..2061 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 2512..3306 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2566..2585) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3176..3195 /label=Neo-F /note="Neomycin resistance gene, forward primer" regulatory 3507..3516 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3513..3887 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" rep_origin 4025..4613 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4514..4533 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4767..4784 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat 241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcag 301 tgtgagggag tgtaaagctg gtttgatctg gcctccgcgc cgggttttgg cgcctcccgc 361 gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc 421 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga 481 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt 541 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga 601 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca 661 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg 721 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc 781 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg 841 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga 901 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg 961 gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt attcgggtga 1021 gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc 1081 ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta 1141 cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg 1201 cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg 1261 gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg 1321 gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga 1381 agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg 1441 caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag actagtaaat 1501 tgtccgctaa attctggccg tttttggctt ttttgttaga cgaagctaac gcgctaggac 1561 ccgacattag cgctacagct taagcgggat ccactagtaa cggccgccag tgtgctggaa 1621 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg gcccaattcg 1681 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa 1741 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 1801 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct atacgtacgg 1861 cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag 1921 agtgatatta ttgacacgcc ggggcgacgg atggtgatcc ccctggccag tgcacgtctg 1981 ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg 2041 cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct 2101 gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata 2161 taaatgtcag gcatgagatt atcaaaaagg atcttcacct agatcctttt cacgtagaaa 2221 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca 2281 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag 2341 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct 2401 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tctcgccgcc aaggatctga 2461 tggcgcaggg gatcaagctc tgatcaagag acaggatgag gatcgtttcg catgattgaa 2521 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 2581 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 2641 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag 2701 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 2761 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 2821 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 2881 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 2941 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3001 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat 3061 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3121 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3181 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3241 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3301 ttctgaatta ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc 3361 ggtatttcac accgcataca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 3421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3481 tgcttcaata atagcacgtg aggagggcca ccatggccaa gttgaccagt gccgttccgg 3541 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct 3601 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca 3661 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg 3721 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct 3781 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg 3841 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctaaaac 3901 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 3961 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 4021 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 4081 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 4141 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 4201 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 4261 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 4321 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 4381 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 4441 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 4501 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 4561 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 4621 gcaacgcggc ctttttacgg ttcctgggct tttgctggcc ttttgctcac atgttctttc 4681 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 4741 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag //