LOCUS Exported 7261 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSYN-PACE-001 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7261) AUTHORS Peters RT, Toby G, Lu Q, Liu T, Kulman JD, Low SC, Bitonti AJ, Pierce GF TITLE Biochemical and functional characterization of a recombinant monomeric Factor VIII-Fc fusion protein. JOURNAL J Thromb Haemost. 2012 Dec 4. doi: 10.1111/jth.12076. PUBMED 23205847 REFERENCE 2 (bases 1 to 7261) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7261 /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 2217..2226 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3125..3166 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 3176..3193 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3216..3233) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3222..3446 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3492..3920 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3579..3598) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3789..3810 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3929..3949) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3934..4264 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4115..4250 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4177..4196 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 4312..4359 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4378..4776 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 4934..5055 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4971..4990) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5025..5044 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5104..5120) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5104..5120) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5117..5139) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5128..5144 /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(5152..5182) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5197..5218 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5335..5352) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5506..6094) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5586..5605) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6265..7125) /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 6888..6907 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7126..7230) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gtttaaactt aagcttgcca tggagctgag gccctggttg ctatgggtgg tagcagcaac 961 aggaaccttg gtcctgctag cagctgatgc tcagggccag aaggtcttca ccaacacgtg 1021 ggctgtgcgc atccctggag gcccagcggt ggccaacagt gtggcacgga agcatgggtt 1081 cctcaacctg ggccagatct tcggggacta ttaccacttc tggcatcgag gagtgacgaa 1141 gcggtccctg tcgcctcacc gcccgcggca cagccggctg cagagggagc ctcaagtaca 1201 gtggctggaa cagcaggtgg caaagcgacg gactaaacgg gacgtgtacc aggagcccac 1261 agaccccaag tttcctcagc agtggtacct gtctggtgtc actcagcggg acctgaatgt 1321 gaaggcggcc tgggcgcagg gctacacagg gcacggcatt gtggtctcca ttctggacga 1381 tggcatcgag aagaaccacc cggacttggc aggcaattat gatcctgggg ccagttttga 1441 tgtcaatgac caggaccctg acccccagcc tcggtacaca cagatgaatg acaacaggca 1501 cggcacacgg tgtgcggggg aagtggctgc ggtggccaac aacggtgtct gtggtgtagg 1561 tgtggcctac aacgcccgca ttggaggggt gcgcatgctg gatggcgagg tgacagatgc 1621 agtggaggca cgctcgctgg gcctgaaccc caaccacatc cacatctaca gtgccagctg 1681 gggccccgag gatgacggca agacagtgga tgggccagcc cgcctcgccg aggaggcctt 1741 cttccgtggg gttagccagg gccgaggggg gctgggctcc atctttgtct gggcctcggg 1801 gaacgggggc cgggaacatg acagctgcaa ctgcgacggc tacaccaaca gtatctacac 1861 gctgtccatc agcagcgcca cgcagtttgg caacgtgccg tggtacagcg aggcctgctc 1921 gtccacactg gccacgacct acagcagtgg caaccagaat gagaagcaga tcgtgacgac 1981 tgacttgcgg cagaagtgca cggagtctca cacgggcacc tcagcctctg cccccttagc 2041 agccggcatc attgctctca ccctggaggc caataagaac ctcacatggc gggacatgca 2101 acacctggtg gtacagacct cgaagccagc ccacctcaat gccaacgact gggccaccaa 2161 tggtgtgggc cggaaagtga gccactcata tggctacggg cttttggacg caggcgccat 2221 ggtggccctg gcccagaatt ggaccacagt ggccccccag cggaagtgca tcatcgacat 2281 cctcaccgag cccaaagaca tcgggaaacg gctcgaggtg cggaagaccg tgaccgcgtg 2341 cctgggcgag cccaaccaca tcactcggct ggagcacgct caggcgcggc tcaccctgtc 2401 ctataatcgc cgtggcgacc tggccatcca cctggtcagc cccatgggca cccgctccac 2461 cctgctggca gccaggccac atgactactc cgcagatggg tttaatgact gggccttcat 2521 gacaactcat tcctgggatg aggatccctc tggcgagtgg gtcctagaga ttgaaaacac 2581 cagcgaagcc aacaactatg ggacgctgac caagttcacc ctcgtactct atggcaccgc 2641 ccctgagggg ctgcccgtac ctccagaaag cagtggctgc aagaccctca cgtccagtca 2701 ggcctgtgtg gtgtgcgagg aaggcttctc cctgcaccag aagagctgtg tccagcactg 2761 ccctccaggc ttcgcccccc aagtcctcga tacgcactat agcaccgaga atgacgtgga 2821 gaccatccgg gccagcgtct gcgccccctg ccacgcctca tgtgccacat gccaggggcc 2881 ggccctgaca gactgcctca gctgccccag ccacgcctcc ttggaccctg tggagcagac 2941 ttgctcccgg caaagccaga gcagccgaga gtccccgcca cagcagcagc cacctcggct 3001 gcccccggag gtggaggcgg ggcaacggct gcgggcaggg ctgctgccct cacacctgcc 3061 tgagtgagaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggccctt 3121 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca 3181 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 3241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 3301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 3361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 3421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 3481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 3541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 3601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg 3661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 3721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 3781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 3841 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta 3901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 3961 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 4021 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 4081 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 4141 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 4201 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 4261 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat 4321 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg 4381 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac 4441 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc 4501 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg 4561 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat 4621 gagaacaggg gcatcttgag cccctgcgga cggtgtcgac aggtgcttct cgatctgcat 4681 cctgggatca aagcgatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt 4741 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga 4801 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 4861 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 4921 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 4981 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 5041 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 5101 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 5161 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 5221 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 5281 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5341 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 5401 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5701 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5941 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 7141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 7261 c //