LOCUS       sequence-258192-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Rab5A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  TITLE     Expresses HA-tagged human Rab5A in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4767)
  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            "Unpublished"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-Rab5A
FEATURES             Location/Qualifiers
     source          1..4767
                     /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(795..814)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(796..814)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    819..953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(843..862)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     897..916
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1066..1086)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1066..1084)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1105..1121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1105..1121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1118..1140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1129..1145
                     /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(1153..1159,1160..1177,1178..1183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1198..1219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1336..1353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1507..2095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1587..1606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2266..3057,3058..3126))
                     /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     2889..2908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3127..3231)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3257..3712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3394..3415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3606..3625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3783..4767
                     /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 atccatggct agtcgaggcg caacaagacc caacgggcca aatactggaa ataaaatatg
      181 ccagttcaaa ctagtacttc tgggagagtc cgctgttggc aaatcaagcc tagtgcttcg
      241 ttttgtgaaa ggccaatttc atgaatttca agagagtacc attggggctg cttttctaac
      301 ccaaactgta tgtcttgatg acactacagt aaagtttgaa atatgggata cagctggtca
      361 agaacgatac catagcctag caccaatgta ctacagagga gcacaagcag ccatagttgt
      421 atatgatatc acaaatgagg agtcctttgc aagagcaaaa aattgggtta aagaacttca
      481 gaggcaagca agtcctaaca ttgtaatagc tttatcggga aacaaggccg acctagcaaa
      541 taaaagagca gtagatttcc aggaagcaca gtcctatgca gatgacaata gtttattatt
      601 catggagaca tccgctaaaa catcaatgaa tgtaaatgaa atattcatgg caatagctaa
      661 aaaattgcca aagaatgaac cacaaaatcc aggagcaaat tctgccagag gaagaggagt
      721 agaccttacc gaacccacac aaccaaccag gaatcagtgt tgtagtaact aactcgagcc
      781 tctagattct gcagccctat agtgagtcgt attacgtaga tccagacatg ataagataca
      841 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      901 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      961 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     1021 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     1081 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1141 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     1201 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1261 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1321 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1381 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1441 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1921 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2161 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2221 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2341 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2461 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2701 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2761 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2941 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3121 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3181 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3241 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3301 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3361 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3421 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3481 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3541 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3601 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3661 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     3721 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3781 cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     3841 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     3901 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc
     3961 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     4021 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     4081 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     4141 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     4201 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     4261 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     4321 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     4381 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     4441 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     4501 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     4561 gtacattact attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg
     4621 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     4681 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     4741 tatataagca atgctcgttt agggaac
//
