LOCUS       sequence-353252-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in insect cells. Entire CDS, M1-N635. Codon 
            optimised..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6834)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: SYKA-c014
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4044..4061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4071..4241
                     /codon_start=1
                     /product="Z domain of staphylococcal protein A, engineered 
                     to contain multiple positive charges (Hedhammar and Hober, 
                     2007)"
                     /label=Z-basic tag
                     /note="useful for purifying tagged proteins by 
                     cation-exchange chromatography"
                     /translation="VDNKFNKERRRARREIRHLPNLNREQRRAFIRSLRDDPSQSANLL
                     AEAKKLNDAQPK"
     CDS             4248..4268
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    6324..6458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6361..6380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6415..6434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6487..6652
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatcgggcg cggatccatg ggacatcacc atcatcacca ctcaagcggg gttgacaata
     4081 aattcaacaa ggagcgacgt cgggcacgcc gtgaaatacg acacctccct aatctcaatc
     4141 gagaacaacg gcgtgcgttt ataaggagtc tgcgtgacga ccctagccaa tcagctaact
     4201 tgttggctga agccaagaaa cttaacgatg ctcaacccaa aggcacggag aatttgtatt
     4261 tccagtcgat ggcatcctct ggaatggcag attctgccaa tcacctaccc tttttcttcg
     4321 gcaatataac tagagaggaa gcagaggatt accttgtgca gggtgggatg tcggacggac
     4381 tctacctgct tcgccaatcg agaaactacc taggcgggtt cgccctgtca gtggcccatg
     4441 gtagaaaggc gcaccactat acgatcgaga gggagctcaa cggaacctac gctatcgcgg
     4501 gtggccggac tcatgcgagc ccggctgatt tatgtcatta ccatagtcaa gagagcgatg
     4561 gattggtctg tttattgaag aaacctttca accgccctca gggcgtgcaa ccgaaaaccg
     4621 ggccctttga ggaccttaag gaaaacctca tacgtgagta cgtaaagcag acttggaact
     4681 tgcaagggca ggcactagag caagcgatta tttcccagaa gcctcaacta gaaaaactga
     4741 ttgcaaccac ggcacatgaa aaaatgcctt ggttccatgg gaaaatctcc cgagaagaat
     4801 cagaacaaat agttttaatt gggtcgaaaa caaacggcaa gtttctcatt cgggctaggg
     4861 ataataatgg gagttacgcc ctgtgtcttc tgcatgaggg aaaggtctta cactaccgca
     4921 tcgataaaga taaaaccggg aagttatcaa tcccagaagg caaaaaattc gatactctat
     4981 ggcagttagt cgagcactac tcatacaagg ctgatggact cctgcgggta ttgacagttc
     5041 cgtgccaaaa gatcggcaca cagggaaacg tgaattttgg aggtcggccc cagttaccag
     5101 gttcccatcc agcgacctgg agcgcaggag ggataatatc gagaatcaaa tcctattcct
     5161 ttccgaagcc aggtcaccga aaatcgtctc ccgctcaagg gaacaggcaa gaatctacag
     5221 tcagtttcaa cccatacgag cccgagctgg cgccctgggc cgcggataaa ggcccgcagc
     5281 gtgaggcatt gcccatggac accgaggtgt acgaatctcc ctatgctgat cctgaggaga
     5341 ttagaccaaa ggaagtctac cttgaccgga agctactgac attggaggac aaagaactcg
     5401 gtagtggaaa ttttggtacg gtaaaaaagg gctattacca gatgaagaag gtggttaaaa
     5461 cggttgctgt aaaaatccta aagaacgaag ccaacgaccc agcccttaag gacgaactgc
     5521 tagctgaagc gaatgtgatg cagcagctag ataatcctta tatagtaaga atgatcggaa
     5581 tatgtgaggc cgaaagttgg atgctagtaa tggagatggc ggaattggga cctctcaata
     5641 aatatcttca acagaacagg catgtaaaag acaaaaatat cattgaattg gtgcaccagg
     5701 tgtctatggg gatgaaatac cttgaagagt caaacttcgt acatcgcgat ctagcagccc
     5761 gaaatgtcct cctggttacg caacactatg ccaagatatc ggatttcggt cttagcaagg
     5821 cattaagggc agatgaaaat tattataagg ctcaaactca cgggaaatgg ccggttaagt
     5881 ggtatgcgcc agagtgcatt aattactata agttttcgag caagagcgac gtttggagtt
     5941 ttggtgtcct catgtgggaa gctttttcct atggccagaa gccttaccgc ggaatgaaag
     6001 gttctgaggt aactgcgatg ttagaaaaag gcgagcgtat gggctgtccg gctggttgcc
     6061 cgcgtgaaat gtatgacttg atgaatcttt gctggactta tgatgtcgag aacaggccag
     6121 gttttgcagc cgtggaactg cgcttaagaa actactatta tgacgttgtt aattgactcg
     6181 aggcatgcgg taccaagctt gtcgagaagt actagaggat cataatcagc cataccacat
     6241 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6301 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6361 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6421 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     6481 gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     6541 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     6601 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     6661 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     6721 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     6781 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga atgg
//
