LOCUS       Exported                6512 bp DNA     linear   SYN 28-OCT-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6512)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6512
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1076..2035
                     /organism="Mus musculus"
                     /chromosome="16"
                     /strain="C57BL/6J"
                     /mol_type="mRNA"
                     /note="color: #ffffff"
                     /db_xref="taxon:10090"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     769..789
                     /label=CMV Forward
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     primer_bind     863..882
                     /label=T7_F
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(933..952)
                     /label=PCDNA Linear Reverse
                     /note="color: black; sequence: GCTGTTGCACCTCCTCATgg"
     CDS             935..1027
                     /codon_start=1
                     /label=QSOX1 SignalP
                     /note="color: #a6acb3"
                     /translation="MRRCNSGSGPPPSLLLLLLWLLAVPGANAAP"
     CDS             1034..1051
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     misc_feature    1052..1072
                     /label=TEV site
                     /note="color: #00ff00"
     primer_bind     1054..1097
                     /label=OLI390 ST6 F His Tev
                     /note="color: black; sequence: 
                     GAATCTCTATTTCCAGGGAGGTAAGCCACAGCCCTCCTTGCAGG; added: 
                     2020-11-30"
     CDS             1070..1075
                     /label=G-G-Linker
                     /note="color: #00ffff; direction: RIGHT"
     gene            1076..2035
                     /gene="St6gal1"
                     /gene_synonym="AW742324; Si; Siat1; St6G; St6gal;St6Gal-I; 
                     St6galI"
                     /label=St6gal1
                     /note="beta galactoside alpha 2,6 sialyltransferase 
                     1;Derived by automated computational analysis using gene 
                     prediction method: Gnomon. Supporting evidence includes 
                     similarity to: 3 mRNAs, 31 ESTs, 349 long SRA reads, 13 
                     Proteins, and 100% coverage of the annotated genomic 
                     feature by RNAseq alignments, including 27 samples with 
                     support for all annotated introns"
                     /note="color: #a6acb3; direction: RIGHT"
                     /db_xref="GeneID:20440"
                     /db_xref="MGI:MGI:108470"
     primer_bind     1076..1102
                     /label=ST6GAL1 f2
                     /note="color: black; sequence: AAGCCACAGCCCTCCTTGCAGGTGTGG;
                     added: 2020-11-30"
     primer_bind     1226..1264
                     /label=OLI410 Fwd C139R
                     /note="color: black; sequence: 
                     GAGGCGCTGCGCGCCCACCTTCGAGACCACGTGAATGTG; added: 2021-06-21"
     primer_bind     complement(1856..1927)
                     /label=OLI411 Rev C350R and C361R
                     /note="color: black; sequence: 
                     CGGATGGTAGGCACCCATCGTGGCGGCGCTGTCAAAGAACTTCTGGTGATAGTAGGCCA
                     CATCTGTCTTGCG; added: 2021-06-21"
     primer_bind     complement(2006..2035)
                     /label=ST6GAL1 r2
                     /note="color: black; sequence: 
                     TCAACAGCGATTGTTCCGGAAGCCAGACAG; added: 2020-11-30"
     primer_bind     complement(2012..2057)
                     /label=OLI391 ST6 R His Tev
                     /note="color: black; sequence: 
                     GTGCTGGATATCTGCAGAATTCTCAACAGCGATTGTTCCGGAAGCC; added: 
                     2020-11-30"
     primer_bind     complement(2012..2057)
                     /label=OLI413 ST6 R C403R
                     /note="color: black; sequence: 
                     GTGCTGGATATCTGCAGAATTCTCAgcgGCGATTGTTCCGGAAGCC; added: 
                     2021-06-21"
     primer_bind     complement(2012..2029)
                     /label=OLI412 Rev C403R (Debbie)
                     /note="color: black; sequence: 
                     CTCAATGATGGTGATGATGGTGTCAAGCGCGATTGTTCCGGAAGCC; added: 
                     2021-06-21"
     primer_bind     complement(2106..2128)
                     /label=OLI27_BGH_R_72C
                     /note="color: black; sequence: gcaactagaaggcacagtcgagg"
     primer_bind     complement(2106..2123)
                     /label=BGH rev
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    2112..2223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      2382..2810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2824..3153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3004..3139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     complement(3066..3085)
                     /label=sv40-rev
                     /note="color: black; sequence: CCTCGGCCTCTGCATAAATA"
     CDS             3220..4014
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4188..4309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4225..4244)
                     /label=SV40 pa-R
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     complement(4358..4374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4382..4398
                     /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)."
                     /note="color: #31849b"
     promoter        complement(4406..4436)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4406 .. 4412 / #ffffff / -10
                        2: 4413 .. 4430 / #ffffff
                        3: 4431 .. 4436 / #ffffff / -35"
     protein_bind    4451..4472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4760..5345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5516..6376)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5516 .. 6307 / #ccffcc
                        2: 6308 .. 6376 / #ccffcc / signal sequence
                     Cleavage site after base 6307"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6377..6481)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
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 gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccatgagg aggtgcaaca gcggctccgg
      961 gccgccgccg tcgctgctgc tgctgctgct gtggctgctc gcggttcccg gcgctaacgc
     1021 ggccccgcag ggtcatcatc atcaccatca cgagaatctc tatttccagg gaggtaagcc
     1081 acagccctcc ttgcaggtgt gggacaagga ctccacatac tcaaaactta accccaggct
     1141 gctgaagatc tggaggaact atctgaacat gaataaatat aaagtgtcct acaaggggcc
     1201 gggaccagga gtcaagttca gcgtagaggc gctgcgctgc caccttcgag accacgtgaa
     1261 tgtgtctatg atagaggcca cagattttcc cttcaacacc actgaatggg agggttacct
     1321 gcccaaggag aacttcagaa ccaaggctgg gccttggcat aagtgtgccg tcgtgtcttc
     1381 tgcaggatct ctgaagaact cccagctggg tcgagagatt gataatcatg atgcggtcct
     1441 gaggtttaat ggggcaccta cagacaactt ccaacaggat gtgggcacaa aaactaccat
     1501 ccgcctagtg aactctcagt tagtcaccac agaaaagcgc ttcctgaagg acagtttgta
     1561 caccgaagga atcctgattc tgtgggaccc atctgtgtat catgcagaca ttccgcagtg
     1621 gtatcagaag ccagactaca acttcttcga aacctataag agttaccgaa ggcttcaccc
     1681 cagccagcct ttttacatcc tcaagcccca gatgccatgg gaactatggg acatcattca
     1741 ggaaatctct ccagatctga ttcagccgaa tcccccatcc tccggcatgc tgggtatcat
     1801 cattatgatg acgctgtgtg accaagttga tatttacgag ttcctcccat ccaagcgcaa
     1861 gacagatgtg tgctactatc accagaagtt ctttgacagc gcctgcacga tgggtgccta
     1921 ccatccgctc ctcttcgaga agaatatggt gaagcatctc aatgagggaa cagatgaaga
     1981 catttatttg tttgggaaag ctaccctgtc tggcttccgg aacaatcgct gttgagaatt
     2041 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
     2101 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2161 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2221 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2281 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2341 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2401 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2461 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2521 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2581 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2641 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2701 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2761 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2821 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2881 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3061 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3121 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3181 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3241 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3301 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3361 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3421 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3481 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3541 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3601 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3661 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3721 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3781 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3841 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3901 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3961 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4021 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5281 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5341 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5401 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5461 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5521 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5581 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5641 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5701 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5761 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5821 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5881 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5941 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6001 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6061 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6121 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6181 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6241 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6301 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6361 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6421 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6481 gcacatttcc ccgaaaagtg ccacctgacg tc
//
