LOCUS       sequence-230371-        5502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Rabin8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5502)
  TITLE     HA-Rabin8
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5502)
  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            "Unpublished"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-Rabin8
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1070..1096
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2508..2527)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2509..2527)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2532..2666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2556..2575)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2610..2629
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2779..2799)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2779..2797)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2818..2834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2818..2834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2831..2853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2842..2858
                     /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(2866..2872,2873..2890,2891..2896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2911..2932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3049..3066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3220..3808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3300..3319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3979..4770,4771..4839))
                     /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     4602..4621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4840..4944)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4970..5425)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5107..5128)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5319..5338
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat accatgcgat acccatatga
     1081 tgtgccagat tacgctggat ccatggctaa tgatcccttg gaaggcttcc atgaagtaaa
     1141 ccttgcttca cctacttctc cggaccttct tggtgtgtat gaatcaggaa ctcaagagca
     1201 gactacctca ccaagtgtca tctaccggcc acacccttca gctttatcct ctgtacctat
     1261 ccaggcaaat gcattagatg tttctgaact tcctacacaa cccgtgtatt catcccccag
     1321 acgtttaaat tgtgcggaaa tatctagtat cagctttcat gttacagacc cagccccttg
     1381 ctctacctct ggagtcacag ctggattaac taaattaact acaagaaagg acaactataa
     1441 tgcagagaga gagtttttac agggtgctac tataacagag gcttgcgatg gcagtgatga
     1501 tatttttggg ttgagtactg atagtctgtc tcgtttacga agcccatctg ttttggaagt
     1561 tagagaaaag ggctatgaac gattaaaaga agaactcgca aaagctcaga gggaactgaa
     1621 gttaaaagat gaagaatgtg agaggctttc aaaagtgcga gatcaacttg gacaggaatt
     1681 ggaagaactc acagctagtc tatttgagga agctcataaa atggtgagag aagcaaatat
     1741 caagcaggca acagcagaaa aacagctaaa agaagcacaa ggaaaaattg atgtacttca
     1801 agctgaagta gctgcattga agacacttgt attgtccagt tctccaacat cacctacgca
     1861 ggagcctttg ccaggtggaa agacaccttt taaaaagggg catacaagaa ataaaagcac
     1921 aagcagtgct atgagtggca gtcatcagga cctcagtgtg atacagccaa ttgtaaaaga
     1981 ctgcaaagag gctgacttat ccttgtataa tgaattccga ttgtggaagg atgagcccac
     2041 aatggacagg acgtgtcctt tcttagacaa aatctaccag gaagatatct ttccatgttt
     2101 aacattctca aaaagtgagt tggcttcagc tgttctggag gctgtggaaa acaatactct
     2161 aagcattgaa ccagtgggat tacaacctat ccggtttgtg aaagcttctg cagttgaatg
     2221 cggaggacca aaaaaatgtg ctctcactgg ccagagtaag tcctgtaaac acagaattaa
     2281 attaggggac tcaagcaact attattatat ttctcctttt tgcagataca ggatcacttc
     2341 tgtatgtaac ttttttacat acattcgata cattcagcag ggactcgtga aacagcagga
     2401 tgttgatcag atgttttggg aggttatgca gttgagaaaa gagatgtcat tggcaaagct
     2461 gggttatttc aaagaggaac tctgactcga gcctctagat tctgcagccc tatagtgagt
     2521 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
     2581 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2641 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2701 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc
     2761 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     2821 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2881 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2941 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3061 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3661 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4861 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4921 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     4981 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5041 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5101 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5161 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5221 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5281 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5341 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5401 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     5461 gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga cc
//
