LOCUS       sequence-176728-        3748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used for in situ hybridisation or other purposes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3748)
  AUTHORS   Hall TE, Martel N, Lo HP, Xiong Z, Parton RG
  TITLE     A plasmid library of full-length zebrafish rab proteins for in vivo 
            cell biology.
  JOURNAL   Cell Logist. 2017 Mar 1;7(1):e1301151. doi: 
            10.1080/21592799.2017.1301151. eCollection 2017.
  PUBMED    28396820
REFERENCE   2  (bases 1 to 3748)
  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: "Cell 
            Logist."; date: "2017-03-1"; volume: "7(1):e1301151. doi"; pages: " 
            10.1080/21592799.2017.1301151. eCollection 2017"
COMMENT     Sequence Label: Rab33ba cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..200
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     192..209
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     193..209
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     216..235
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        216..234
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      265..274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      269..278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1105..1123)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1106..1123)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1141..1157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1141..1157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1154..1176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1165..1181
                     /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(1189..1195,1196..1213,1214..1219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1234..1255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1372..1389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1543..2131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1623..1642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2302..3093,3094..3162))
                     /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     2925..2944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3163..3267)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3345..3748,1..52))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3482..3503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3694..3713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
       61 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
      121 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
      181 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
      241 tgggcccgac gtcgcatgct cccggccgcc atggcggccg cgggaattcg attgcatggc
      301 agatatcgag tcctcttttg agttcagctc ttctctgacc agctcgtcac tgcctcctcc
      361 tcggacacgc atctttaaaa tcatcgtgat tggagactca ggggttggca agacttgcct
      421 cacctataga ttctgtgcag gcaagtttcc agacaagact gaggccacta ttggggtgga
      481 tttccgagag aaggttattg agatagatgg cgagaaaatc aaggtccagt tgtgggacac
      541 ggcaggccag gagcgtttcc gtaagagcat ggtgcagcac tactaccgca acgtccatgc
      601 tgtggtgttt gtctatgacg tcaccaatgc cgccagcttc cgaagcctcc cagcgtggat
      661 tgaagagtgt cgccaacatg cactcggtca ggaggtgcca agaatccttg tcggcaacaa
      721 atgcgaccta cgtcacgccg ctcaagtaag cacagatgtt gcacagcagt ttgcagatac
      781 acactcaatg cctctttttg agacctcggc caaaaacccc tacgggaatg aagacggcac
      841 tcagaataat agtgaccacg tggaggccat ttttatgaca gtggcgcaca agctgaaatc
      901 ccaaaagccg ctggttttaa gccagccacc ttgcggatat ggagatactg tcactcttag
      961 gaggcaagac caggaggacg ggggaaactg gggctgcggt tgttggagaa gctaatgaag
     1021 tgcaaatcac tagtgaattc gcggccgcct gcaggtcgac catatgggag agctcccaac
     1081 gcgttggatg catagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg
     1141 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1201 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1321 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1981 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2581 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3181 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3241 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt
     3301 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     3361 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3421 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3481 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3541 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3601 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3661 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3721 tagggcgctg gcaagtgtag cggtcacg
//
