LOCUS       sequence-258201-        5227 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses HA-tagged human IST1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  TITLE     Expresses HA-tagged human IST1 in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5227)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HA-IST1
FEATURES             Location/Qualifiers
     source          1..5227
                     /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(1255..1274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1256..1274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1279..1413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1303..1322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1357..1376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1526..1546)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1526..1544)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1565..1581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1565..1581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1578..1600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1589..1605
                     /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(1613..1619,1620..1637,1638..1643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1658..1679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1796..1813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1967..2555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2726..3517,3518..3586))
                     /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     3349..3368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3587..3691)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3717..4172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4066..4085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4243..5227
                     /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 atccaaggcc tctcgaggca tgctgggctc tggatttaaa gctgagcgct taagagtgaa
      181 tttgagatta gtcataaatc gccttaaact attggagaaa aagaaaacgg aactggccca
      241 gaaagcaagg aaggagattg ctgactatct ggctgctggg aaagatgaac gagctcggat
      301 ccgtgtggag cacattatcc gggaagacta cctcgtggag gccatggaga tcctggagct
      361 gtactgtgac ctgctgctgg ctcggtttgg ccttatccag tctatgaagg aactagattc
      421 tggtctggct gaatctgtgt ctacattgat ctgggctgct cctcgactcc agtcagaagt
      481 ggctgagttg aaaatagttg ctgatcagct ctgtgccaag tatagcaagg aatatggcaa
      541 gctatgtagg accaaccaga ttggaactgt gaatgacagg ctaatgcaca agctgagtgt
      601 ggaagcccca cccaaaatcc tggtggagag atacctgatt gaaattgcaa agaattacaa
      661 cgtaccctat gaacctgact ctgtggtcat ggcagaagct cctcctgggg tagagacaga
      721 tcttattgat gttggattca cagatgatgt gaagaaagga ggccctggaa gaggagggag
      781 tggtggcttc acagcaccag ttggtggacc tgatggaacg gtgccaatgc ccatgcccat
      841 gcccatgcct atgccatctg caaatacgcc tttctcatat ccactgccaa agggaccatc
      901 agatttcaat ggactgccaa tggggactta tcaggccttt cccaatattc atccacctca
      961 gataccagca actcccccat cgtatgaatc tgtagatgac attaatgctg ataagaatat
     1021 ctcttctgca cagattgttg gtcctggacc caagccagaa gcctctgcaa agcttccttc
     1081 cagacctgca gataactatg acaactttgt cctaccagag ttgccatctg tgccagacac
     1141 actaccaact gcatctgctg gtgccagcac ctcagcatct gaagacattg actttgatga
     1201 tctttcccgg aggtttgaag agctgaaaaa gaaaacatag tctagattct gcagccctat
     1261 agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     1321 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1381 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1441 tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat
     1501 tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     1561 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1621 gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1681 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1741 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1801 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1861 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2401 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2761 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2821 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2881 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2941 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3001 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3061 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3121 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3181 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3241 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3301 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3361 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3421 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3481 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3541 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3601 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3661 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     3721 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3781 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3841 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3901 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3961 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4021 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4081 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4141 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     4201 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat
     4261 atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat
     4321 atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat
     4381 atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg
     4441 aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg
     4501 ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa
     4561 tatataggca atatccaata tggccctatg ccaatatggc tattggccag gttcaatact
     4621 atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag
     4681 cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc
     4741 actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc
     4801 ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc
     4861 tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc
     4921 attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca
     4981 cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact attggaagta
     5041 cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg
     5101 ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc
     5161 attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt
     5221 agggaac
//
