LOCUS Exported 7712 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION human Soluble Flt1 isoform e15a (terminates in exon 15a). ACCESSION . VERSION . KEYWORDS sFlt1-e15a V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7712) AUTHORS Thomas CP, Andrews JI, Raikwar NS, Kelley EA, Herse F, Dechend R, Golos TG, Liu KZ TITLE A recently evolved novel trophoblast-enriched secreted form of fms-like tyrosine kinase-1 variant is up-regulated in hypoxia and preeclampsia. JOURNAL J Clin Endocrinol Metab. 2009 Jul;94(7):2524-30. doi: 10.1210/jc.2009-0017. Epub 2009 Mar 31. PUBMED 19336504 REFERENCE 2 (bases 1 to 7712) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7712 /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 3210..3251 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 3261..3278 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3301..3318) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3307..3531 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3577..4005 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3664..3683) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3874..3895 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4014..4034) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4019..4348 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4199..4334 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4261..4280 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4415..5209 /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(4469..4488) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5079..5098 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5385..5506 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5422..5441) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5476..5495 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5555..5571) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5555..5571) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5568..5590) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5579..5595 /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(5603..5633) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5648..5669 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5786..5803) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5957..6545) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6037..6056) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6716..7576) /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 7339..7358 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7577..7681) /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 gctggctagt 901 taagcttggt accgagctcg gatccagtac ccttcaccat ggtcagctac tgggacaccg 961 gggtcctgct gtgcgcgctg ctcagctgtc tgcttctcac aggatctagt tcaggttcaa 1021 aattaaaaga tcctgaactg agtttaaaag gcacccagca catcatgcaa gcaggccaga 1081 cactgcatct ccaatgcagg ggggaagcag cccataaatg gtctttgcct gaaatggtga 1141 gtaaggaaag cgaaaggctg agcataacta aatctgcctg tggaagaaat ggcaaacaat 1201 tctgcagtac tttaaccttg aacacagctc aagcaaacca cactggcttc tacagctgca 1261 aatatctagc tgtacctact tcaaagaaga aggaaacaga atctgcaatc tatatattta 1321 ttagtgatac aggtagacct ttcgtagaga tgtacagtga aatccccgaa attatacaca 1381 tgactgaagg aagggagctc gtcattccct gccgggttac gtcacctaac atcactgtta 1441 ctttaaaaaa gtttccactt gacactttga tccctgatgg aaaacgcata atctgggaca 1501 gtagaaaggg cttcatcata tcaaatgcaa cgtacaaaga aatagggctt ctgacctgtg 1561 aagcaacagt caatgggcat ttgtataaga caaactatct cacacatcga caaaccaata 1621 caatcataga tgtccaaata agcacaccac gcccagtcaa attacttaga ggccatactc 1681 ttgtcctcaa ttgtactgct accactccct tgaacacgag agttcaaatg acctggagtt 1741 accctgatga aaaaaataag agagcttccg taaggcgacg aattgaccaa agcaattccc 1801 atgccaacat attctacagt gttcttacta ttgacaaaat gcagaacaaa gacaaaggac 1861 tttatacttg tcgtgtaagg agtggaccat cattcaaatc tgttaacacc tcagtgcata 1921 tatatgataa agcattcatc actgtgaaac atcgaaaaca gcaggtgctt gaaaccgtag 1981 ctggcaagcg gtcttaccgg ctctctatga aagtgaaggc atttccctcg ccggaagttg 2041 tatggttaaa agatgggtta cctgcgactg agaaatctgc tcgctatttg actcgtggct 2101 actcgttaat tatcaaggac gtaactgaag aggatgcagg gaattataca atcttgctga 2161 gcataaaaca gtcaaatgtg tttaaaaacc tcactgccac tctaattgtc aatgtgaaac 2221 cccagattta cgaaaaggcc gtgtcatcgt ttccagaccc ggctctctac ccactgggca 2281 gcagacaaat cctgacttgt accgcatatg gtatccctca acctacaatc aagtggttct 2341 ggcacccctg taaccataat cattccgaag caaggtgtga cttttgttcc aataatgaag 2401 agtcctttat cctggatgct gacagcaaca tgggaaacag aattgagagc atcactcagc 2461 gcatggcaat aatagaagga aagaataaga tggctagcac cttggttgtg gctgactcta 2521 gaatttctgg aatctacatt tgcatagctt ccaataaagt tgggactgtg ggaagaaaca 2581 taagctttta tatcacagat gtgccaaatg ggtttcatgt taacttggaa aaaatgccga 2641 cggaaggaga ggacctgaaa ctgtcttgca cagttaacaa gttcttatac agagacgtta 2701 cttggatttt actgcggaca gttaataaca gaacaatgca ctacagtatt agcaagcaaa 2761 aaatggccat cactaaggag cactccatca ctcttaatct taccatcatg aatgtttccc 2821 tgcaagattc aggcacctat gcctgcagag ccaggaatgt atacacaggg gaagaaatcc 2881 tccagaagaa agaaattaca atcagagatc aggaagcacc atacctcctg cgaaacctca 2941 gtgatcacac agtggccatc agcagttcca ccactttaga ctgtcatgct aatggtgtcc 3001 ccgagcctca gatcacttgg tttaaaaaca accacaaaat acaacaagag cctgaactgt 3061 atacatcaac gtcaccatcg tcatcgtcat catcaccatt gtcatcatca tcatcatcgt 3121 catcatcatc atcatcaaag ggtcaagaca attctgcaga tatccagcac agtggcggcc 3181 gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc 3241 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag 3301 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 3361 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 3421 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 3481 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 3541 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 3601 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 3661 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 3721 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 3781 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 3841 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 3901 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 3961 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 4021 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 4081 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 4141 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 4201 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 4261 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 4321 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 4381 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 4441 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 4501 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 4561 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 4621 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 4681 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 4741 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 4801 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 4861 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 4921 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 4981 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 5041 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 5101 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 5161 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 5221 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 5281 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 5341 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 5401 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5461 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 5521 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 5581 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 5641 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 5701 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 5761 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 5821 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 5881 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 5941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 6001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 6061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 6121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 6181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 6241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 6301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 6361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 6421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 6481 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 6541 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 6601 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 6661 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 6721 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 6781 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 6841 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 6901 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 6961 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 7021 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7081 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7141 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7201 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7261 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 7321 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 7381 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 7441 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 7501 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 7561 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 7621 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 7681 gcacatttcc ccgaaaagtg ccacctgacg tc //