LOCUS       sequence-473025-        5196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Septin12 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5196)
  TITLE     expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5196)
  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: 
            "Unpublished"
COMMENT     Sequence Label: HA-SEPT12
FEATURES             Location/Qualifiers
     source          1..5196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             92..118
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1224..1243)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1225..1243)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1248..1382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1272..1291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1326..1345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1495..1515)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1495..1513)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1534..1550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1534..1550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1547..1569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1558..1574
                     /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(1582..1588,1589..1606,1607..1612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1627..1648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1765..1782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1936..2524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2016..2035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2695..3486,3487..3555))
                     /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     3318..3337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3556..3660)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3686..4141)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3823..3844)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4035..4054
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4212..5196
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg ataccatgcg atacccatat gatgtgccag attacgctgg
      121 atctatggac cccctgaggc gctccccctc tccctgcctg tcctcgcagc cctccagccc
      181 cagcacccca ccctgcgaga tgcttggtcc tgtgggcatt gaggccgtgc tggaccagct
      241 gaagatcaag gctatgaaga tggggtttga gttcaacatc atggtggtgg ggcaaagcgg
      301 gctgggcaag tccacgatgg tgaacacgct gttcaagtcc aaagtgtgga agtcaaaccc
      361 accgggcttg ggggtgccca caccccagac gctgcagctg cattcactga cccatgtcat
      421 agaggagaag ggtgtgaagc tgaagctgac ggtgacggac acgcccggct tcggggacca
      481 gatcaacaat gacaactgct gggaccccat cctgggctac atcaacgagc aatacgagca
      541 gtacctgcag gaggagatcc tcatcacccg ccagcgccac atcccagaca cccgggtaca
      601 ctgctgcgtg tactttgtac cacccactgg gcactgcctg cggcccctgg acattgagtt
      661 cctgcagcgg ctgtgccgga ctgtgaatgt ggtgcccgtg attgccaggg ccgacagcct
      721 gaccatggag gagcgagagg ccttcaggcg caggatccag cagaacctga ggacccactg
      781 catcgacgtc tacccccaga tgtgctttga cgaggacatc aatgacaaaa tcctcaacag
      841 caagttacgg gaccgaatcc cttttgccgt ggtaggggct gaccaagagc acctgatgaa
      901 cgggaggtgt gtcctgggcc ggaagaccaa gtggggcatc attgaagtgg agaacatggc
      961 gcactgtgaa tttcctctcc tgagagacct gcttatccgc tcccacctcc aagacctgaa
     1021 ggacataacc cacaacatcc actatgagaa ctaccgcgtc atcagactca atgaaagcca
     1081 cctgctgccc cgcgggcccg gctgggtgaa cctggccccg gcctccccag gacagctgac
     1141 caccccccgg accttcaagg tctgcagggg ggcccatgac gattctgatg atgagttctg
     1201 actcgagcct ctagattctg cagccctata gtgagtcgta ttacgtagat ccagacatga
     1261 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1321 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1381 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1441 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1501 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1561 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     1621 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1681 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1741 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1801 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1861 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1921 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1981 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2041 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2101 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2161 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2221 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2281 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2341 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2401 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2461 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2521 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2581 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2641 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2701 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2761 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2821 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2881 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2941 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3001 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3061 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3121 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3181 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3241 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3301 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3361 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3421 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3481 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3541 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3601 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3661 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     3721 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3781 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3841 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3901 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3961 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4021 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4081 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4141 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4201 ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata
     4261 tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata
     4321 tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata
     4381 tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga
     4441 ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg
     4501 cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc
     4561 caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca
     4621 tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata
     4681 tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat
     4741 atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca
     4801 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
     4861 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
     4921 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
     4981 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
     5041 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
     5101 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
     5161 tgggaggtct atataagcaa tgctcgttta gggaac
//
