LOCUS       sequence-483011-        7350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Fc-fusion pST3Gal1 H302A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7350)
  AUTHORS   Hombu R, Beatty LE, Tomaszewski J, Park S, Neelamegham S
  TITLE     Engineering glycosyltransferases into glycan binding proteins using 
            a mammalian surface display platform.
  JOURNAL   Nat Commun. 2025 Jul 18;16(1):6637. doi: 10.1038/s41467-025-62018-z.
  PUBMED    40681527
REFERENCE   2  (bases 1 to 7350)
  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: "Nat 
            Commun."; date: "2025-07-18"; volume: "16(1):6637. doi"; pages: " 
            10.1038/s41467-025-62018-z"
COMMENT     Sequence Label: pCSCG-Fc-pST3Gal1 H302A
FEATURES             Location/Qualifiers
     source          1..7350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             928..945
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1015..1029
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             2601..2621
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2622..2639
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     LTR             3152..3332
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3358..3375)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3364..3588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3634..4062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3721..3740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3931..3952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4071..4091)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4076..4405
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4256..4391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4318..4337
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4453..4500
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4519..4893
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5023..5144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5060..5079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5114..5133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5193..5209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5193..5209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5206..5228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5217..5233
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5241..5247,5248..5265,5266..5271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5286..5307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5424..5441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5595..6183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5675..5694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6354..7145,7146..7214))
                     /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     6977..6996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7215..7319)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctagcgccg ccaccatgat tcctgccaga tttgccgggg tgctgcttgc tctggccctc
      901 attttgccag ggaccctttg taccggtcac catcatcatc atcatcaggc caccgaatat
      961 gagtacctag attatgattt cctgccagaa acggagcctc cacgacctat gatggatgac
     1021 gatgacaaat ctagaacatg cccaccgtgc ccagcacctg aactcgcggg ggcaccgtca
     1081 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc
     1141 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg
     1201 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg
     1261 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac
     1321 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc
     1381 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc
     1441 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg
     1501 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac
     1561 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag
     1621 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag
     1681 agcctctccc tgtctccggg taaaggtagc ggctctggta gcggttccgg ctctcgcccg
     1741 tgcacctgta cccgctgcat cgaggagcag cgcgttagcg cgtggtttga tgaacgtttc
     1801 aatcgcagca tgcagccgct gttaaccgcg aaaaatgcgc atctggaaga agatacctat
     1861 aaatggtggt tacgcctgca gcgtgagaaa cagccaaaca acctgaatga tacgatccgc
     1921 gagctcttcc aagtggtgcc gggcaacgtt gatccgctgc tggaaaaacg cttagtgtct
     1981 tgtcgccgct gcgccgtggt gggcaactct ggtaacctga aagaaagcta ctatggcccg
     2041 cagattgaca gccatgactt cgttctgcgt atgaacaaag cgccgaccga aggttttgag
     2101 gcggatgttg gtagcaaaac cacgcatcat tttgtttatc cggaaagctt ccgtgaatta
     2161 gcgcaagaag tgagcatgat tctggtaccg tttaaaacga ccgatctgga atgggtgatt
     2221 tctgccacga ccacgggccg tatttctcat acgtatgttc cggtgccggc gaaaatcaaa
     2281 gtgaagaaag aaaaaattct gatctatcac ccggcgttta ttaaatatgt gtttgatcgc
     2341 tggctgcagg gccacggtcg ttatccgtct acgggcatcc tgagcgtgat ttttagcctg
     2401 catatttgtg acgaggtcga cctgtacggt tttggcgccg attctaaagg taactggcac
     2461 gcgtattggg aaaacaatcc atctgcgggt gcgtttcgca agaccggcgt tcatgatggt
     2521 gatttcgagt ctaatgttac caccatcctg gcgagcatta acaaaattcg tattttcaaa
     2581 ggtcgttgac ctgcaggttc gaaaacctgt acttccaggg ccatcaccat caccatcact
     2641 gactcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     2701 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     2761 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     2821 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     2881 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     2941 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     3001 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     3061 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     3121 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     3181 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     3241 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     3301 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
     3361 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3421 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3481 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3541 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3601 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3661 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3721 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3781 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3841 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3901 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3961 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4021 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4081 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4141 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4201 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4261 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4321 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4381 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4441 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     4501 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     4561 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     4621 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     4681 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     4741 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     4801 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     4861 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     4921 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4981 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5041 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5101 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5161 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5221 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5281 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5341 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5401 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5461 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5521 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5581 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5641 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5701 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5761 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5821 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5881 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5941 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6001 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6061 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6121 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6181 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6241 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6301 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6361 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6421 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6481 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6541 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6601 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6661 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6721 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6781 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6841 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6901 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6961 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7021 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7081 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7141 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7201 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7261 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7321 acatttcccc gaaaagtgcc acctgacgtc
//
