LOCUS       sequence-322801-        5138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SBP tagged partial length ST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5138)
  AUTHORS   Sun X, Mahajan D, Chen B, Song Z, Lu L
  TITLE     A quantitative study of the Golgi retention of glycosyltransferases.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272560. doi: 
            10.1242/jcs.258564. Epub 2021 Oct 21.
  PUBMED    34533190
REFERENCE   2  (bases 1 to 5138)
  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: "J Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272560. doi"; 
            pages: " 10.1242/jcs.258564. Epub 2021 Oct 21"
COMMENT     Sequence Label: SBP-ST*-GFP
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             625..738
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     regulatory      1099..1108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1105..1107,1108..1110,1111..1824)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1150..1171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1946..2067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1983..2002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2037..2056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2074..2529)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2211..2232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2423..2442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2556..2660
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2662..3019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2662..2853
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2685..2705)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2870..3005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3054..3848
                     /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             3054..3848
                     /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(3108..3127)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3718..3737
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4036..4055)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4080..4127
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4456..5044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4945..4964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc caccatggac gagaagacca ctggttggcg aggtggacac
      661 gttgttgaag gactggctgg ggaacttgaa caacttcgtg cacgactgga gcatcaccca
      721 caaggtcaac gtgaaccact cgaggccacc atgattcaca ccaacctgaa gaaaaagttc
      781 agctgctgcg tactagtctt tcttctgttt gcagtcatat gtgtgtggaa ggaaaagaag
      841 aaagggagtt actatgattc ctttaaattg caaaccaagg aattccaggt gttaaagagt
      901 ctggggaaat tggccatggg gtctgattcc cagtctgtat cctcaagcag cacccaggac
      961 ccccacaggg gccgccagac cctcggcagt ctcagaggcc tagccaaggc caaaccagag
     1021 gcctccttcc aggtgtggaa caaggacagc tcttccaaaa accttatccc taggctgcaa
     1081 aagatctggg caccggtcgc caccatggtg tccaagggcg aggagctgtt caccggggtg
     1141 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttctc cgtgcggggc
     1201 gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatcag caccaccggc
     1261 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagagcttc
     1321 tcccgctacc ccgaccacat gaagcgccac gacttcttca agagcgccat gcccgaaggc
     1381 tacgtccagg agcgcaccat ctccttcaag gacgacggca cctacaagac ccgcgccgag
     1441 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1501 gaggacggca acatcctggg gcacaagctg gagtacaact tcaactccca caacgtctat
     1561 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg
     1621 gaggacggct ccgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1681 cccgtgctgc tgcccgacaa ccactacctg tccacccagt ccaagctgtc caaagacccc
     1741 aacgagaagc gcgatcacat ggtccttctg gaattcgtga ccgccgccgg gatcactcac
     1801 ggcatggacg agctgtacaa gtaagcggcc gcgactctag atcataatca gccataccac
     1861 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     1921 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1981 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2041 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2101 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2161 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2221 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2281 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2341 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2401 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2461 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2521 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2581 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2641 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2701 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2761 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2821 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2881 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2941 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3001 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3061 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3121 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3181 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3241 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3301 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3361 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3421 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3481 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3541 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     3601 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3661 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3721 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3781 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3841 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3901 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3961 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4021 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4081 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4141 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4201 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4261 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4321 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4381 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4441 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4501 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4561 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4621 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4681 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4741 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4801 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4861 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4921 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4981 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5041 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5101 catgttcttt cctgcgttat cccctgattc tgtggata
//
