LOCUS       sequence-226349-        5247 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of ARL4a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5247)
  TITLE     Kahn expression plasmids
REFERENCE   2  (bases 1 to 5247)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: rk4s
FEATURES             Location/Qualifiers
     source          1..5247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..88
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        69..87
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    115..139
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(764..788)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(841..859)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      855..902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1268..1273,1274..1290,1291..1296))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1323..1341)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1409..1513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1514..1582,1583..2374)
                     /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(1732..1751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2545..3133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3034..3053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3287..3304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3319..3458
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3384..3403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3544..3566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3560..3751)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5213..5232
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
       61 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gatcacaagt
      121 ttgtacaaaa aagcaggctt agaaggagat agaaccatgg ggaatgggct gtcagaccag
      181 acttctatcc tgtccaacct gccttcattt cagtctttcc acattgttat tctgggtttg
      241 gactgtgctg gaaagacaac tgtcttatac aggctgcagt tcaatgaatt tgtaaatacc
      301 gtacctacca aaggatttaa cactgagaaa attaaggtaa ccttgggaaa ttctaaaaca
      361 gtcacttttc acttctggga tgtaggtggt caggagaaat taaggccact gtggaagtca
      421 tataccagat gcacagatgg cattgtattt gttgtggact ctgttgatgt cgaaaggatg
      481 gaagaagcca aaactgaact tcacaaaata actaggatat cagaaaatca gggagtccct
      541 gtacttatag ttgctaacaa acaagatttg aggaactcat tgtcactttc agaaattgag
      601 aaattgttag caatgggtga actgagctca tcaactcctt ggcatttgca gcctacctgt
      661 gcaatcatag gagatggcct aaaggaagga cttgagaaac tacatgatat gatcattaaa
      721 agaagaaaaa tgttgcggca acagaaaaag aaaagatgat aggacccagc tttcttgtac
      781 aaagtggtga tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      841 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      901 tgctgaaagg aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatcgcg
      961 tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg
     1021 gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc
     1081 acgccatagt gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt
     1141 accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg
     1201 agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa
     1261 ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa
     1321 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1381 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1441 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1501 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1561 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1621 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1681 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1741 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1801 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1861 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1921 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1981 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2041 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2101 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2161 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2221 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2281 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2341 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2401 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2461 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2521 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2581 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2641 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2701 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2761 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2821 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2881 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2941 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3001 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3061 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3121 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3181 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3241 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3301 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3361 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3421 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3481 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3541 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3601 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3661 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3721 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     3781 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     3841 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3901 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     3961 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     4021 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     4081 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     4141 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     4201 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc
     4261 tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc
     4321 tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc
     4381 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
     4441 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
     4501 ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc
     4561 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     4621 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     4681 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
     4741 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     4801 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     4861 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     4921 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     4981 gactgggttg aaggctctca agggcatcgg tcgatcgacg ctctccctta tgcgactcct
     5041 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     5101 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     5161 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     5221 cggcgatata ggcgccagca accgcac
//
