LOCUS       sequence-258194-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Rab7A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  TITLE     Expresses HA-tagged human Rab7A in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4743)
  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            "Unpublished"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-Rab7A
FEATURES             Location/Qualifiers
     source          1..4743
                     /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(771..790)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(772..790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    795..929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(819..838)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     873..892
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1042..1062)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1042..1060)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1081..1097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1081..1097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1094..1116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1105..1121
                     /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(1129..1135,1136..1153,1154..1159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1174..1195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1312..1329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1483..2071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1563..1582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2242..3033,3034..3102))
                     /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     2865..2884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3103..3207)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3233..3688)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3370..3391)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3582..3601
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3759..4743
                     /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 atccatgacc tctaggaaga aagtgttgct gaaggttatc atcctgggag attctggagt
      181 cgggaagaca tcactcatga accagtatgt gaataagaaa ttcagcaatc agtacaaagc
      241 cacaatagga gctgactttc tgaccaagga ggtgatggtg gatgacaggc tagtcacaat
      301 gcagatatgg gacacagcag gacaggaacg gttccagtct ctcggtgtgg ccttctacag
      361 aggtgcagac tgctgcgttc tggtatttga tgtgactgcc cccaacacat tcaaaaccct
      421 agatagctgg agagatgagt ttctcatcca ggccagtccc cgagatcctg aaaacttccc
      481 atttgttgtg ttgggaaaca agattgacct cgaaaacaga caagtggcca caaagcgggc
      541 acaggcctgg tgctacagca aaaacaacat tccctacttt gagaccagtg ccaaggaggc
      601 catcaacgtg gagcaggcgt tccagacgat tgcacggaat gcacttaagc aggaaacgga
      661 ggtggagctg tacaacgaat ttcctgaacc tatcaaactg gacaagaatg accgggccaa
      721 ggcctcggca gaaagctgca gttgctgact cgagcctcta gattctgcag ccctatagtg
      781 agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
      841 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      901 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      961 aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg
     1021 cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     1081 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1141 cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     3241 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3301 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3361 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3421 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3481 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3541 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3601 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3661 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     3721 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
     3781 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
     3841 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
     3901 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
     3961 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
     4021 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
     4081 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
     4141 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
     4201 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
     4261 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
     4321 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
     4381 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
     4441 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
     4501 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
     4561 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     4621 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     4681 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     4741 aac
//
