LOCUS       sequence-226665-        5250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ARL4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5250)
  TITLE     Kahn expression plasmids
REFERENCE   2  (bases 1 to 5250)
  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"
COMMENT     Sequence Label: rk16s
FEATURES             Location/Qualifiers
     source          1..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     130..149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        130..148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    176..200
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(828..852)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(905..923)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      919..966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1332..1337,1338..1354,1355..1360))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1387..1405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1473..1577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1578..1646,1647..2438)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1796..1815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2609..3197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3098..3117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3351..3368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3383..3522
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3448..3467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3608..3630
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3624..3815)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
       61 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
      121 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agatcacaag
      181 tttgtacaaa aaagcaggct tagaaggaga tagaaccatg gggaaccact tgactgagat
      241 ggcgcccact gcctcctcct tcttgcccca cttccaagcc ctgcatgtcg tggtcattgg
      301 gctggactct gctggaaaga cctccctcct ttaccgcctc aagttcaagg agtttgtcca
      361 gagtgtcccc accaaaggct tcaacaccga gaagatccgg gtgcccctcg ggggatcgcg
      421 tggcatcacc ttccaagtgt gggacgtcgg ggggcaggag aagctgcgac cactgtggcg
      481 ctcttatacc cgccggacag acggtctagt gtttgtggtg gacgctgcgg aggctgagcg
      541 gctggaggaa gccaaggtgg agttgcaccg aatcagccgg gcctcggaca accagggcgt
      601 gccagtgctg gtgctggcca acaagcagga ccagcccggg gcactgagcg ctgctgaggt
      661 ggagaagagg ctggcagtcc gagagctagc agccgccact ctcactcatg tgcaaggctg
      721 cagcgctgtg gacggtctgg gcctgcagca gggccttgag cgcctctatg agatgatcct
      781 caagaggaag aaggcagctc ggggtggcaa gaagagacgg tgataggacc cagctttctt
      841 gtacaaagtg gtgatgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
      901 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      961 tttttgctga aaggaggaac tatatccgga tatccacagg acgggtgtgg tcgccatgat
     1021 cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg
     1081 gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag
     1141 cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca agaggcccgg
     1201 cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac
     1261 gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga
     1321 taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac
     1381 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1441 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1501 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1561 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1621 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1681 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1741 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1801 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     1861 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1921 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1981 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2041 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2101 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2161 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2221 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2281 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2341 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2401 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2461 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2521 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3241 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3301 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3361 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3421 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3481 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     3541 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3601 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3661 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     3721 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     3781 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     3841 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     3901 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     3961 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4021 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4081 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4141 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4201 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4261 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
     4321 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
     4381 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     4441 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     4501 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     4561 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     4621 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     4681 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     4741 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     4801 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     4861 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     4921 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     4981 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     5041 agctgactgg gttgaaggct ctcaagggca tcggtcgatc gacgctctcc cttatgcgac
     5101 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     5161 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5221 cccacgccga aacaagcgct catgagcccg
//
