LOCUS       sequence-401590-        6549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing an antigen targeting TRIP8b (exon 1b).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6549)
  TITLE     Trimmer lab antigen plasmids
REFERENCE   2  (bases 1 to 6549)
  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"
COMMENT     Sequence Label: TRIP8b (1b-2)-GFP/pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..6549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..965
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             171..965
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(225..244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     835..854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1153..1172)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1197..1244
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1573..2161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2062..2081
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2336..2639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2640..2843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2794..2814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2882..2891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2888..2890,2891..2893,2894..3604)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2888..2890,2891..2893,2894..3604)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2933..2954)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3194..3213)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3541..3562
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5612..5733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5649..5668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5703..5722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5740..6195)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5877..5898)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6089..6108
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6222..6326
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6328..6549,1..136)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6328..6519
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6351..6371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6536..6549,1..122)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
       61 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
      121 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
      181 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
      241 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
      301 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
      361 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
      421 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
      481 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
      541 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
      601 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
      661 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
      721 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
      781 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
      841 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
      901 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
      961 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     1021 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     1081 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     1141 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     1201 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     1261 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     1321 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     1381 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     1441 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1501 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1561 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1621 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1681 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1741 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1801 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1861 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1921 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1981 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2041 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2101 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2161 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2221 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     2281 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2341 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2401 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2461 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2521 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2581 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2641 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     2701 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     2761 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     2821 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt
     2881 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2941 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3001 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3061 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3121 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3181 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3241 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3301 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3361 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3421 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3481 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3541 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3601 caagtccgga ctcagatctc gagctcaagc ttcgaattct gggatgtctg acagtgaaat
     3661 ggatggaagg actcacattc catctctact taatgccctt ttgtccagaa atcgggtcat
     3721 gcagatgagc tacttgaaaa gtaaagaaca aggctatggc aagctaagca gtgacgagga
     3781 cctggaaata attgttgacc agaagcagct ggtgaatgag caacaagaga gcagacccct
     3841 cctgagcccc tccatcgacg acttcctctg cgaaaccaaa tcggaggcca ttgcaaagcc
     3901 agtgacatcc aacacagccg tgttgaccac tggcttggat ctccttgacc tgagtgaacc
     3961 cgtctctcaa acccaaacca aagccaagaa atctgagccc tcatcaaaga gctcgtccct
     4021 caagaagaaa gctgatggct ccgacctcat cagtgcagat gctgagcaga gagcacaggc
     4081 tctccgaggg ccagagacat catccttaga tttagacatt caaacacaac tggaaaaatg
     4141 ggatgatgtc aagtttcacg gtgaccgaac aagcaagggg catctcatgg cagaaaggaa
     4201 atcatgctct tccagaactg ggtccaaaga gcttctttgg tccgcagaac acagatctca
     4261 gccagagctg agtactggga aaagcgccct caactctgag tcggcttccg agttggaact
     4321 tgtagctcca gcacaggctc ggctgaccaa agagcatcgc tggggaagtg ccctgctctc
     4381 caggaaccat tccttggaag aggagtttga aagggcaaaa gcggcagtag agtcagatac
     4441 agagttctgg gataagatgc aggcagaatg ggaggaaatg gcccggagga actggatatc
     4501 cgagaaccag gaagctcaga accaagtcac cgtctcggcc agtgagaagg gatattactt
     4561 ccacactgaa aatcccttca aggactggcc tggagcattt gaagaaggct taaaaaggtt
     4621 gaaggaaggg gacctgcctg tcaccatcct gttcatggaa gctgcaattc ttcaggaccc
     4681 tggagacgct gaggcatggc agtttcttgg tataacccag gctgagaatg aaaatgagca
     4741 ggcagctatt gtcgccctcc agaggtgcct ggagttgcag cccaacaact tgaaggctct
     4801 gatggccttg gctgtaagtt acaccaacac cagccaccag caggatgctt gtgaggcact
     4861 gaagaactgg attaagcaaa acccaaagta caagtacctc gtgaagaaca agaaaggctc
     4921 tccaggcctc acacggcgga tgtctaaatc cccagttgac agctctgttc tggaaggagt
     4981 gaaggagttg tacctggaag ctgcccacca aaatggtgac atgattgacc cagacctgca
     5041 gacaggtctg ggagttctgt tccacctgag tggagagttc aatagagcaa tcgatgcatt
     5101 taatgctgcc ttaacagttc ggccagagga ttactcactg tggaaccgtc ttggggcaac
     5161 cttggcaaat ggagaccgca gtgaggaagc tgtggaggcc tacacgagag cactggagat
     5221 ccagccaggt ttcatccgct ccagatacaa cctggggatc agctgcatca acctgggtgc
     5281 ctacagagaa gcggtcagca acttcctcac tgccctcagc ttgcagagga agagcaggaa
     5341 ccagcagcaa gtccctcacc ctgccatctc cgggaacatc tgggccgccc tcaggattgc
     5401 cctgtcgctg atggaccagc cggagctctt ccaggccgcc aacctcggtg acttggatgt
     5461 cctcctgaga gctttcaatt tggatccttg aaggtctaga taactgatca taatcagcca
     5521 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     5581 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5701 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     5761 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5821 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5881 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5941 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     6001 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     6061 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     6121 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     6181 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6241 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6301 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     6361 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6421 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6481 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6541 gcccctaac
//
