LOCUS       sequence-176737-        3704 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 3704)
  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 3704)
  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: Rab27a cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     192..214
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     206..223
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     207..223
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     230..249
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        230..248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      279..288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      283..292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1075..1093)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1076..1093)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1111..1127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1111..1127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1124..1146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1135..1151
                     /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(1159..1165,1166..1183,1184..1189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1204..1225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1342..1359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1513..2101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1593..1612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2272..3063,3064..3132))
                     /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     2895..2914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3133..3237)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3315..3704,1..66))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3452..3473)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3664..3683
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
       61 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      121 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      181 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc
      241 actatagggc gaattgggcc cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa
      301 ttcgattggt ctcatcatgt ccgatgggga ctatgattac ctcatcaagt tcctcgctct
      361 gggggactct ggggtgggaa agaccagctt cctatatcag tacacagaca gcaaattcaa
      421 ctccaaattc ataaccacag tgggaataga cttcagagaa aaacgagtgg tgtataaatc
      481 cagtggtcct gatggcacag caggcagagg ccagaggatc cacatccagc tttgggatac
      541 ggccggacag gagaggtttc gaagtttgac cactgctttc ttcagagatg cgatgggctt
      601 tctgctgctc tttgacctca ccaacgagca gagcttcctg aatgtgcgca actggatgag
      661 tcagctgcag acacatgcgt actgtgagaa tcctgatatc gtcttgtgtg gaaataaatc
      721 agacctggag gagcagcgtg ccgtcaaagc ggagaacgca agagaactgg cagagaaata
      781 cggcgtgcca tatttcgaga cgagcgcggc gaacggtgag aatgtgagtc gtgcggtgga
      841 ggttctgctg gatctgatca tgaagcgcat ggagagatgt gtggataaat cctggatccc
      901 cgacgggaca gtccgcagca acggccacag caccgccgac ctgacagaac cacacgaccc
      961 cgagcagagc aagtgtgcat gctaataatc agcgaatcac tagtgaattc gcggccgcct
     1021 gcaggtcgac catatgggag agctcccaac gcgttggatg catagcttga gtattctata
     1081 gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1141 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1201 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1261 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1321 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1381 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1441 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1501 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1561 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1621 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1681 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1741 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1801 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1861 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1921 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1981 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2041 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2101 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2161 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2221 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2281 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2341 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2401 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2461 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2521 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2581 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2641 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2701 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2761 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2821 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2881 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2941 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3001 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3061 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3121 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3181 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3241 atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3301 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3361 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3421 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3481 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3541 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3601 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3661 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaa
//
