LOCUS Exported 7040 bp ds-DNA circular SYN 23-SEP-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Luciferase-pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7040) AUTHORS Safran M, Kim WY, O'Connell F, Flippin L, Gunzler V, Horner JW, Depinho RA, Kaelin WG TITLE Mouse model for noninvasive imaging of HIF prolyl hydroxylase activity: assessment of an oral agent that stimulates erythropoietin production. JOURNAL Proc Natl Acad Sci U S A. 2006 Jan 3. 103(1):105-10. PUBMED 16373502 REFERENCE 2 (bases 1 to 7040) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7040 /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" regulatory 918..927 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 924..2576 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(990..1008) /label=LucNrev /note="Firefly luciferase, reverse primer" promoter complement(2593..2611) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2594..2611) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2631..2648) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2637..2861 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2907..3335 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2994..3013) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3204..3225 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3344..3364) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3349..3678 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3529..3664 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3591..3610 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3745..4539 /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(3799..3818) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4409..4428 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4713..4834 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4750..4769) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4804..4823 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4883..4899) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4883..4899) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4896..4918) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4907..4923 /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(4931..4961) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4976..4997 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5114..5131) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5285..5873) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5365..5384) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6044..6904) /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 6667..6686 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6905..7009) /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 gcttggcatt 901 ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa ggcccggcgc 961 cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat 1021 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt 1081 acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga 1141 atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt 1201 tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg 1261 aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt 1321 tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg 1381 attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac 1441 ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg 1501 cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata 1561 gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc 1621 cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac 1681 tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt 1741 ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct 1801 ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg 1861 cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc 1921 tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac 1981 ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg 2041 ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga 2101 gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg 2161 acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct 2221 tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg 2281 aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc 2341 ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga 2401 tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc 2461 gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa 2521 gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaattct 2581 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt 2641 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2701 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 2761 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 2821 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 2881 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 2941 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3001 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3061 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3121 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3181 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3241 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3301 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3361 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3421 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3601 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3661 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 3721 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3781 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3841 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3901 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 3961 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4021 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4081 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4141 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4201 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4261 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4321 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4381 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4441 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4501 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4561 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4621 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4681 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4741 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4801 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4861 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4921 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 4981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 7021 gaaaagtgcc acctgacgtc //