LOCUS       sequence-405387-        6108 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression of human Syntaxin 4 without transmembrane helix
            and a C-terminal cysteine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6108)
  TITLE     Mammalian expression of GFP-tagged Lamin-A
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6108)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: hSTX4(1-271,272C)
FEATURES             Location/Qualifiers
     source          1..6108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(17..64)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     60..78
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(131..148)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(977..994)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1004..1021)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1040..1062
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1077..1101
                     /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)."
     primer_bind     complement(1101..1120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1102..1120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1204..1223)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1429..1506
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1507..2589
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1526..1545)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3398..3589
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3583..3605)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3691..3830
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3746..3765
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3845..3862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4016..4604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4096..4115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4715..4734)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4726..5541
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4726..5541
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4799..4818)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5633..6088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5770..5791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5982..6001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
       61 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
      121 ccggatctca gtggtggtgg tggtggtgct cgagctagca tttacgagct tttttctggt
      181 tttccagagc ggttttaacg tgttcctgac cacgttcaac gtagtcagca gaagacagga
      241 tgtttttttc gatacggttg atcatttcac cctgcatttc aacttcggta gccaggaagg
      301 tgaagatgtc gtgcagttca cggatagaac gttccagctg ctggatttca gagtgacgag
      361 cagagatttc gttcagagcc tgacgggtaa cctgggtgtc tttcaggatg ttagaaacga
      421 aaacttcaga ctgaccagag tccagcatct gttccagttc ttcgtcagaa accataccag
      481 cgttggtgat tttcagctga cgacggatac gttcaacgtt tttttcacgg tattcagact
      541 gcatagagtt gcatttgttg atcagttcaa cgaactgctg agacagaaca ccgtgctggg
      601 ttttacgcat acgggtgtta acagagttgt agttttcgtc agcttcttct ttctgcggtt
      661 cgatagcttt cagctgcaga cggatttcac gacccagctg tttgatttcg tcacgcaggt
      721 tctgcagttc ctgtttcata gattcttccg gcagcggggt agccaggatg gtaacctgct
      781 gtttttccag ttcctgaact ttgttaccca gtttaacgat ggtctgacgg atggtacgaa
      841 ctttgtggaa gaattcttcg tccggagaac ccagacgagc ggtacccggg tgaacaacca
      901 gagcaacacg ttctttgtct tcttcgtcag aagagtcgtc accctgacgc agttcgtggg
      961 tacggtcacg catatggctg ccgcgcggca ccaggccgct gctgtgatga tgatgatgat
     1021 ggctgctgcc catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
     1081 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct
     1141 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     1201 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     1261 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     1321 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
     1381 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
     1441 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     1501 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     1561 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     1621 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     1681 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     1741 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     1801 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     1861 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     1921 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     1981 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     2041 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     2101 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     2161 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     2221 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     2281 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     2341 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     2401 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     2461 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     2521 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2581 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
     2641 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2701 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2761 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     2821 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     2881 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg
     2941 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     3001 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     3061 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     3121 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     3181 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     3241 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     3301 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     3361 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     3421 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     3481 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     3541 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3601 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3661 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     3721 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3781 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     3841 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3901 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3961 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4021 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4081 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4141 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4201 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4261 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4321 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4381 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4441 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4501 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4561 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4621 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4681 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     4741 cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg
     4801 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     4861 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     4921 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     4981 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca
     5041 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     5101 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     5161 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     5221 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     5281 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     5341 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     5401 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     5461 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     5521 atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta
     5581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     5641 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5701 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5761 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5821 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5881 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5941 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6001 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6061 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccaa tccggata
//
