LOCUS       sequence-503401-        4598 bp    DNA     circular SYN 19-MAR-2026
DEFINITION  Contains a multiple cloning site to insert a gene downstream of 
            Flag-TurboID (N-term aa1-194).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4598)
  TITLE     Trinkle Lab plasmids
REFERENCE   2  (bases 1 to 4598)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFlagTurboNTD(C1)
FEATURES             Location/Qualifiers
     source          1..4598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             122..916
                     /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             122..916
                     /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(176..195)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     786..805
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1104..1123)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1148..1195
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1524..2112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2013..2032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2287..2590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2591..2794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2745..2765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2826..2849
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3436..3488
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3612..3733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3649..3668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3703..3722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3740..4195)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3877..3898)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4089..4108
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4222..4326
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4328..4598,1..87)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4328..4519
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4351..4371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4536..4598,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
       61 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
      121 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      181 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      241 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      301 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      361 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      421 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      481 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
      541 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
      601 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
      661 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
      721 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
      781 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
      841 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
      901 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
      961 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1021 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1081 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     1141 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     1201 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     1261 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     1321 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     1381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1441 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1501 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1561 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1621 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1981 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2041 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2101 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2161 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2221 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2281 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2341 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2401 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2461 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2521 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2581 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2641 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2701 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2761 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     2821 gcatggacta caaagacgat gacgacaagg gctctaaaga caatactgtg cctctgaagc
     2881 tgatcgctct cctggctaat ggcgagttcc atagtggcga acagctggga gaaaccctgg
     2941 gcatgtccag ggccgctatc aacaagcaca ttcagactct gcgcgactgg ggcgtggacg
     3001 tgttcaccgt gcccggaaag ggctactctc tgcccgagcc tatcccgctg ctgaacgcta
     3061 aacagattct gggacagctg gacggcggga gcgtggcagt cctgcctgtg gttgactcca
     3121 ccaatcagta cctgctggat cgaatcggcg agctgaagag tggggatgct tgcattgcag
     3181 aatatcagca ggcagggaga ggaagcagag ggaggaaatg gttctctcct tttggagcta
     3241 acctgtacct gagtatgttt tggcgcctga agcggggacc agcagcaatc ggcctgggcc
     3301 cggtcatcgg aattgtcatg gcagaagcgc tgcgaaagct gggagcagac aaggtgcgag
     3361 tcaaatggcc caatgacctg tatctgcagg atagaaagct ggcaggcatc ctggtggagc
     3421 tggccggaat aacatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc
     3481 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     3541 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3601 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3721 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3781 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3841 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3901 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3961 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4021 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4081 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4141 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4201 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4261 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4321 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4501 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4561 ccgcccattc tccgccccat ggctgactaa tttttttt
//
