LOCUS Exported 6034 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CMV-SEAP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6034) TITLE pLAI-Delta-mls REFERENCE 2 (bases 1 to 6034) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6034 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(35..1552) /codon_start=1 /product="secreted alkaline phosphatase from human placenta" /label=SEAP /note="The natural enzyme has a C-terminal transmembrane sequence that is replaced by a GPI anchor." /translation="MLGPCMLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAA KKLQPAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKT YNVDKHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGK SVGVVTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVIL GGGRKYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLD PSVTHLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHG HHESRAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGL APGKARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGED VAVFARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD" promoter complement(1717..1920) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(1746..1766) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 1921..2300 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 2402..2421 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2533..2552) /label=T7 /note="T7 promoter, forward primer" promoter complement(2534..2552) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2559..2576) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2559..2575) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2568..2590) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2717..3169 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2801..2820) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3011..3032 /label=F1ori-F /note="F1 origin, forward primer" promoter 3195..3299 /gene="bla" /label=AmpR promoter CDS 3300..4160 /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 complement(3518..3537) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4331..4919 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4820..4839 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5073..5090 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5207..5228 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5243..5273 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5281..5297 /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 5286..5308 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5305..5321 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5305..5321 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5342..5360 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 5406..5540 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5430..5449) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5484..5503 /label=SV40pA-R /note="SV40 polyA, reverse primer" ORIGIN 1 acgcggggac cacgggttaa cccgggtgcg cggcgtcggt ggtgccggcg gggggcgcca 61 ggtcgcaggc ggtgtagggc tccaggcagg cggcgaaggc catgacgtgc gctatgaagg 121 tctgctcctg cacgccgtga accaggtgcg cctgcgggcc gcgcgcgaac accgccacgt 181 cctcgcctgc gtgggtctct tcgtccaggg gcactgctga ctgctgccga tactcggggc 241 tcccgctctc gctctcggta acatccggcc gggcgccgtc cttgagcaca tagcctggac 301 cgtttccgta taggaggacc gtgtaggcct tcctgtcccg ggccttgcca ggggccagcc 361 cgaagatgga gctccctcgc agggggtagc ctccgaagga gaagacgtgg gagtggtcgg 421 cagtgacgag gctcagcgtg tcctcctcgc tggtgagctg gcccgccctc tcaatggcgt 481 cgtcgaacat gatcgtctca gtcagtgccc ggtaagccct gctttcatga tgaccatggt 541 cgatgcgacc accctccacg aagaggaaga agccgcgggg gttcctgctc agcaggcgca 601 gggcagcctc tgtcatctcc atcagggagg ggtccagtgt ggagtctcgg tggatctcgt 661 atttcatgtc tccaggctca aagagaccca tgagatgggt cacagacggg tccagggaag 721 cctgcatgag ctcagtgcgg ttccacacat accgggcacc ctggcgcttc gccagccatt 781 cctgcaccag attcttcccg tccagcctgg tcccaccttg gctgtagtca tctgggtact 841 cagggtctgg ggttcccatg cgaaacatgt actttcggcc tccacctagg atcacgtcaa 901 tgtccatgtt ggagatgagc tgcgtagcga tgtcctggca cccctcctgg cgggccgagg 961 caggcacgtc ggcgtccgag taccagttgc ggttcaccgt gtgggcgtag gtgccggctg 1021 gcgaggcgtg ctgcactcgt gtggtggtta ccactcccac tgacttccct gctttcttgg 1081 cccgattcat cacggagatg acctcgttgc cgcgtgtcgt gttgcactgg ttaaagcggg 1141 cggctgcact caagccaatg gtctggaagt tgcccttgac cccgcacagg taggccgtgg 1201 ctgtggctcc actgtctggc acatgtttgt ctacattgta tgtcttggac agagccacat 1261 atgggaagcg gtccatggcc aggggtatct caggccccag tttgtccttc ttctgccctt 1321 ttaggatcct ggcagctgtc accgtagaca cccccatccc atcgcccagg aagatgatga 1381 ggttcttggc ggctgtctgt gcaggctgca gcttcttggc ggcacccagg gcctcggctg 1441 cctcgcggtt ccagaagtcc gggttctcct cctcaactgg gatgatgccc agggagagct 1501 gtagcctcag gcccagcagc agcagcagca gcagcatgca gggccccagc atgtctggag 1561 ggcagcagga cagtggcgag gcaggaatta gcttgaggtg cacaccaatg tggtgaatgg 1621 tcaaatggcg tttattgtat cgagctaggc acttaaatac aatatctctg caatgcggaa 1681 ttcgatatca agcttatcga tacggttcac taaacgagct ctgcttatat agacctccca 1741 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa 1801 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg 1861 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac 1921 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt 1981 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt 2041 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac 2101 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga 2161 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt 2221 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata 2281 actagtcaat aatcaatgtc aacatggcgg taatgttgga catgagccaa tataaatgta 2341 catattatga tatggataca acgtatgcaa tgggccaagc tcatggctga ctaatttttt 2401 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 2461 gcttttttgg aggcctaggc ttttgcaaaa agctcgacgc gaggctcgag ggggggcccg 2521 gtacccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt 2581 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc 2641 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc 2701 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggttacg 2761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 3121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt 3181 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 3241 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 3301 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 3361 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 3421 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 3481 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 3541 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 3601 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 3661 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 3721 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 3781 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc 3841 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 3901 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 3961 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 4021 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 4081 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 4141 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 4201 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 4261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 4321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 4381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 4441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 4501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 4561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 4621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 4681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 4741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 4801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 4861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 4921 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 4981 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 5041 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 5101 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 5161 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 5221 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 5281 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg 5341 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta 5401 ggatcgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 5461 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 5521 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 5581 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt 5641 atgatcaatg aattgctcta gaggatcccc gggtacccac tgggctgtag gtggtggctg 5701 gatggcctga gccccctcct ttcctggggt gcaggcagcc acagggcaca acctgcagtc 5761 cctggggttg gggtagggag tccgagagca gccaaggggc aggtttggtg tcatgggctc 5821 agtccccaga gcgcacgggg aggggaaggt tccatggcag gacctctgta tagggactcc 5881 ggggatgacg actcgggctg gaggccaggc aggacgacag gaggtgggga gcaggagaac 5941 tccgggatgg ggaagcagga gccccaggga cgggacactc agggagcagt ggccgtctcc 6001 agcagcagca gggtcccggc cagcagagga agca //