LOCUS Exported 9599 bp ds-DNA circular SYN 17-AUG-2022 DEFINITION Expression of human HER2 antigen in a Sleeping Beauty transposon vector. ACCESSION . VERSION . KEYWORDS pSBbi G418 Hu Her2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9599) AUTHORS Rad S M AH, Poudel A, Tan GMY, McLellan AD TITLE Promoter choice: Who should drive the CAR in T cells? JOURNAL PLoS One. 2020 Jul 24;15(7):e0232915. doi: 10.1371/journal.pone.0232915. eCollection 2020. PUBMED 32706785 REFERENCE 2 (bases 1 to 9599) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 17, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9599 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(43..60) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 49..273 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter 705..809 /gene="bla" /label=AmpR promoter CDS 810..1670 /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(1028..1047) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1841..2429 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2330..2349 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2583..2600 /label=L4440 /note="L4440 vector, forward primer" primer_bind 2974..2993 /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" polyA_signal 2992..3047 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind 3093..3112 /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" CDS complement(3187..3981) /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(3298..3317) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3908..3927 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter 4614..5795 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" primer_bind 5743..5763 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" ORIGIN 1 aggccaagct tggggatcaa ttctctagag ctcgctgatc agcctcgact gtgccttcta 61 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca 121 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc 181 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata 241 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg 301 cctactagat agcggacccc ttaccgaaac atcgccgcat tctgcagagg agtcgaactt 361 agtgtatgta aacttctgac ccactggaat tgtgatacag tgaattataa gtgaaataat 421 ctgtctgtaa acaattgttg gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac 481 tgacttgcca aaactattgt ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt 541 ttaatgactc caacttaagt gtatgtaaac ttccgacttc aactgtatag ggatccaatt 601 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg 661 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 1021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 1081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 1141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 1201 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 1261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 1321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 1381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 1441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 1501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 1561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 1621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 1681 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 2521 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 2581 gcagcgagtc agtgagcgag gaagcggatc cctatacagt tgaagtcgga agtttacata 2641 caccttagcc aaatacattt aaactcactt tttcacaatt cctgacattt aatcctagta 2701 aaaattccct gtcttaggtc agttaggatc accactttat tttaagaatg tgaaatatca 2761 gaataatagt agagagaatg attcatttca gcttttattt ctttcatcac attcccagtg 2821 ggtcagaagt ttacatacac tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg 2881 tcccgagtag ctagttcatg gcagccagca tatggcatat gttgccaaac tctaaaccaa 2941 atactcattc tgatgtttta aatgatttgc cctcccatat gtccttccga gtgagagaca 3001 caaaaaattc caacacacta ttgcaatgaa aataaatttc ctttattagc cagaagtcag 3061 atgctcaagg ggcttcatga tgtccccata atttttggca gagggaaaaa gatctcagtg 3121 gtatttgtga gccagggcat tggccacacc agccaccacc ttctgatagg cagcctgcac 3181 ctgaggtcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc 3241 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat 3301 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc 3361 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg 3421 ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc 3481 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat 3541 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg 3601 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc 3661 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc 3721 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga 3781 tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa 3841 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt 3901 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg 3961 caatccatct tgttcaatca tggtggcctc aggtgcagag gtttctacag ggaaagagag 4021 aaaggctaca gttagtttgc tgtgatgatg ctaagtgccg aggtcagatg tctatggatt 4081 gtacacggaa tcccaacgca ggaaccaaaa aacactcacc caaaaaaatg gcgctaaggc 4141 cgagagaaag gcgaagttcc gtctacggct atataccttg aaacaggggc ggcagaagtg 4201 acgcaacgcg gttgcgcgga cggcttcctg tgttttcatt ggtcgaagcg ttctgcgcgt 4261 gcgcagagat tccccaggga aggcgggccc tcttccgaat gtccaatccg gacattcttc 4321 cggttggacc tccgcggtca gcgctgtgcc cggcgagaga tcacgtgggg cgcggaggcg 4381 gtgctgctgg ggcacggccg tccagcctcg gcggccatat ttttgagggg ctgttcatct 4441 cgttcacacg ctctgtccgc catgtttgtg agtggaagcg ccattacccc ttcaagcgac 4501 tgaaggctgc agggcctctg gtggcccgca tggggagacc agacccgcca ggcccgcctt 4561 tccgcactca gtccgggctt actttatttt gtgagacagg gtctcgccta gaggctccgg 4621 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 4681 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 4741 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 4801 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg 4861 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac 4921 gcccctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga 4981 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg 5041 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg 5101 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag 5161 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg 5221 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc 5281 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc 5341 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg 5401 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc 5461 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct 5521 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt 5581 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt 5641 ttccccatac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct 5701 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg 5761 ttcaaagttt ttttcttcca tttaaggtgt cgtgaaaact accccaagct ggcctctgag 5821 gccatggagc tggcggcctt gtgccgctgg gggctcctcc tcgccctctt gccccccgga 5881 gccgcgagca cccaagtgtg caccggcaca gacatgaagc tgcggctccc tgccagtccc 5941 gagacccacc tggacatgct ccgccacctc taccagggct gccaggtggt gcagggaaac 6001 ctggaactca cctacctgcc caccaatgcc agcctgtcct tcctgcagga tatccaggag 6061 gtgcagggct acgtgctcat cgctcacaac caagtgaggc aggtcccact gcagaggctg 6121 cggattgtgc gaggcaccca gctctttgag gacaactatg ccctggccgt gctagacaat 6181 ggagacccgc tgaacaatac cacccctgtc acaggggcct ccccaggagg cctgcgggag 6241 ctgcagcttc gaagcctcac agagatcttg aaaggagggg tcttgatcca gcggaacccc 6301 cagctctgct accaggacac gattttgtgg aaggacatct tccacaagaa caaccagctg 6361 gctctcacac tgatagacac caaccgctct cgggcctgcc acccctgttc tccgatgtgt 6421 aagggctccc gctgctgggg agagagttct gaggattgtc agagcctgac gcgcactgtc 6481 tgtgccggtg gctgtgcccg ctgcaagggg ccactgccca ctgactgctg ccatgagcag 6541 tgtgctgccg gctgcacggg ccccaagcac tctgactgcc tggcctgcct ccacttcaac 6601 cacagtggca tctgtgagct gcactgccca gccctggtca cctacaacac agacacgttt 6661 gagtccatgc ccaatcccga gggccggtat acattcggcg ccagctgtgt gactgcctgt 6721 ccctacaact acctttctac ggacgtggga tcctgcaccc tcgtctgccc cctgcacaac 6781 caagaggtga cagcagagga tggaacacag cggtgtgaga agtgcagcaa gccctgtgcc 6841 cgagtgtgct atggtctggg catggagcac ttgcgagagg tgagggcagt taccagtgcc 6901 aatatccagg agtttgctgg ctgcaagaag atctttggga gcctggcatt tctgccggag 6961 agctttgatg gggacccagc ctccaacact gccccgctcc agccagagca gctccaagtg 7021 tttgagactc tggaagagat cacaggttac ctatacatct cagcatggcc ggacagcctg 7081 cctgacctca gcgtcttcca gaacctgcaa gtaatccggg gacgaattct gcacaatggc 7141 gcctactcgc tgaccctgca agggctgggc atcagctggc tggggctgcg ctcactgagg 7201 gaactgggca gtggactggc cctcatccac cataacaccc acctctgctt cgtgcacacg 7261 gtgccctggg accagctctt tcggaacccg caccaagctc tgctccacac tgccaaccgg 7321 ccagaggacg agtgtgtggg cgagggcctg gcctgccacc agctgtgcgc ccgagggcac 7381 tgctggggtc cagggcccac ccagtgtgtc aactgcagcc agttccttcg gggccaggag 7441 tgcgtggagg aatgccgagt actgcagggg ctccccaggg agtatgtgaa tgccaggcac 7501 tgtttgccgt gccaccctga gtgtcagccc cagaatggct cagtgacctg ttttggaccg 7561 gaggctgacc agtgtgtggc ctgtgcccac tataaggacc ctcccttctg cgtggcccgc 7621 tgccccagcg gtgtgaaacc tgacctctcc tacatgccca tctggaagtt tccagatgag 7681 gagggcgcat gccagccttg ccccatcaac tgcacccact cctgtgtgga cctggatgac 7741 aagggctgcc ccgccgagca gagagccagc cctctgacgt ccatcatctc tgcggtggtt 7801 ggcattctgc tggtcgtggt cttgggggtg gtctttggga tcctcatcaa gcgacggcag 7861 cagaagatcc ggaagtacac gatgcggaga ctgctgcagg aaacggagct ggtggagccg 7921 ctgacaccta gcggagcgat gcccaaccag gcgcagatgc ggatcctgaa agagacggag 7981 ctgaggaagg tgaaggtgct tggatctggc gcttttggca cagtctacaa gggcatctgg 8041 atccctgatg gggagaatgt gaaaattcca gtggccatca aagtgttgag ggaaaacaca 8101 tcccccaaag ccaacaaaga aatcttagac gaagcatacg tgatggctgg tgtgggctcc 8161 ccatatgtct cccgccttct gggcatctgc ctgacatcca cggtgcagct ggtgacacag 8221 cttatgccct atggctgcct cttagaccat gtccgggaaa accgcggacg cctgggctcc 8281 caggacctgc tgaactggtg tatgcagatt gccaagggga tgagctacct ggaggatgtg 8341 cggctcgtac acagggactt ggccgctcgg aacgtgctgg tcaagagtcc caaccatgtc 8401 aaaattacag acttcgggct ggctcggctg ctggacattg acgagacaga gtaccatgca 8461 gatgggggca aggtgcccat caagtggatg gcgctggagt ccattctccg ccggcggttc 8521 acccaccaga gtgatgtgtg gagttatggt gtgactgtgt gggagctgat gacttttggg 8581 gccaaacctt acgatgggat cccagcccgg gagatccctg acctgctgga aaagggggag 8641 cggctgcccc agccccccat ctgcaccatt gatgtctaca tgatcatggt caaatgttgg 8701 atgattgact ctgaatgtcg gccaagattc cgggagttgg tgtctgaatt ctcccgcatg 8761 gccagggacc cccagcgctt tgtggtcatc cagaatgagg acttgggccc agccagtccc 8821 ttggacagca ccttctaccg ctcactgctg gaggacgatg acatggggga cctggtggat 8881 gctgaggagt atctggtacc ccagcagggc ttcttctgtc cagaccctgc cccgggcgct 8941 gggggcatgg tccaccacag gcaccgcagc tcatctacca ggagtggcgg tggggacctg 9001 acactagggc tggagccctc tgaagaggag gcccccaggt ctccactggc accctccgaa 9061 ggggctggct ccgatgtatt tgatggtgac ctgggaatgg gggcagccaa ggggctgcaa 9121 agcctcccca cacatgaccc cagccctcta cagcggtaca gtgaggaccc cacagtaccc 9181 ctgccctctg agactgatgg ctacgttgcc cccctgacct gcagccccca gcctgaatat 9241 gtgaaccagc cagatgttcg gccccagccc ccttcgcccc gagagggccc tctgcctgct 9301 gcccgacctg ctggtgccac tctggaaagg cccaagactc tctccccagg gaagaatggg 9361 gtcgtcaaag acgtttttgc ctttgggggt gccgtggaga accccgagta cttgacaccc 9421 cagggaggag ctgcccctca gccccaccct cctcctgcct tcagcccagc cttcgacaac 9481 ctctattact gggaccagga cccaccagag cggggggctc cacccagcac cttcaaaggg 9541 acacctacgg cagagaaccc agagtacctg ggtctggacg tgccagtgtg aggcctgtc //