LOCUS       sequence-68719-e        6281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  General cloning vector for use in E. coli or D. radiodurans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6281)
  AUTHORS   Meima R, Lidstrom ME
  TITLE     Characterization of the minimal replicon of a cryptic Deinococcus 
            radiodurans SARK plasmid and development of versatile Escherichia 
            coli-D. radiodurans shuttle vectors.
  JOURNAL   Appl Environ Microbiol. 2000 Sep;66(9):3856-67.
  PUBMED    10966401
REFERENCE   2  (bases 1 to 6281)
  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: "Appl 
            Environ Microbiol."; date: "2000-09"; volume: "66(9)"; pages: 
            "3856-67"
COMMENT     Sequence Label: pRAD1
FEATURES             Location/Qualifiers
     source          1..6281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(132..200,201..992)
                     /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     complement(350..369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1163..1751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1652..1671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1905..1922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2039..2060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2075..2080,2081..2098,2099..2105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /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)."
     primer_bind     2118..2140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2137..2153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5437..6096
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQSHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5503..5522)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagct cgcgaggcct cgagatctat cgatgcatgc catggtaccc gggagctcga
     2221 attctagaag cttctgcaga cgcgtcgacg tcatatggat ccgattcgac ctgcaggcat
     2281 gcaagctgta cgcctcagct accattcttg cagccgcgtt cagggtcttg acgtgttctt
     2341 tgtatctatc agcaaactga gaacccaatt ctggatcatc cttgtaacgc caactctccg
     2401 accattccga aggaacgcct tgtagtcctc ctgagccgtt agggtatgtc tttagtccaa
     2461 tcaaatggga catttgcaga caggaacgca caagtgtctc acgtcgcttc tctatttcaa
     2521 tatctaaaaa ttcttcactc ttatcgttgc ttgacatatt aaaactgtct aactgatcca
     2581 gttcacttat atggaaagta tcactaaagt catggcttct cagaaattca atacttccca
     2641 ttgatgggga ccggttaacc aaatcgataa aaatttttct gtcatgtttc agataagttt
     2701 ttggcgcgtt gattttataa taaagccgcc attcttgtgc caacaacgcc acgaaaagag
     2761 ccacagcagc agcggccaca tcccattgaa aatctttcgc cagaccaacc gccaggagaa
     2821 ccaagccgaa atagttagcc cagcgaacta aaaaactatc tataaacgta ttaagaaacg
     2881 ttcgggtcat atttccgtga atcatatcag gacgtaaatt tgcacatgac gcatttccgg
     2941 gggcttttat ccctcggact tcttttcagg ccccttctcg gcctttctgt gaccctgttt
     3001 tttcaggccg cgaactgcgc ctggtcgggc ctgaccccca cacgctggcc ttcgtactca
     3061 cgcagcctgc ccagcaggtc ggccagcgcg gcattgacca cggcggcgag gttgcggggc
     3121 ggcgttccgc ctgcggtctg gtcgtgcttc acgtcgcgca gcacgcgcac cagcaccgcg
     3181 cccacgtcgt cggacacgtc ccggccagcg tcggcggcgc gggtgatgtt ccaaatgagt
     3241 ttccgccaga agcccagcga gtcggccccg tccccgaaag ctgccgctag cgcccgcgcc
     3301 tgcctgtcca cgatctcggc gcggtaggcc gggcgggttt cccggtgggc gtcggcaagc
     3361 tcccacacga cgttttcagc gagggagggc gcgggccgga cagtcatgtt atcggagggg
     3421 gagggaagag cggattttat cgcccaggtg ctcacaactt cgtatgtcac ggcaaccggg
     3481 gcgctttttt cgcgcagttc cccgcctgtc acatcctcac agggttcttg actctccggc
     3541 acgctcttag ggagagggtg gagcatgttg tagacggtgc gcccggcctt cgcgtcggcg
     3601 ttcaggtcgc gccagttccg gccccaatcg tcatgcatca ggcggacata tcccgccttc
     3661 ccttccagca cgcgcctcgg cttgagcgtc acggcccaca gcgtcccggt tgccacagac
     3721 tcgccgcgca ggtcgcccat gtgggcatca caggccacca gcccgacccg gcgcagatat
     3781 tgaaggttct cgtagaaggc ggacttcttc agacccacat gcaccatgag cagctcggcg
     3841 ggcaggtgga acacagcgcg ggtgacgtgt tccgcatagc cgcaggcgcg ggccacgtcc
     3901 agcgccaccg agaacagggc gcggaagatg cgccgggccg actcgcggca gggggcgtca
     3961 tctaggcagg cggtgagggt ctgcaccagt tcggcgcgtg tggcgatggt caacggcgcg
     4021 ggcgcgggct gggactgctg ggcgatggtg ggcacgggcg gcggcgagat ggaccgggcc
     4081 gagctggaaa tttcccgtgt gagcgcaggt ttaggcgcat ggttaccttc ctgcagtgaa
     4141 atctgcgaaa tcaggcccgc ttcgaggagt cggaccatga atggattgtg tctcaagaaa
     4201 aaagcctccc ctcttgtcag aggggagggt cccggtctac catgctctca gcgacgagat
     4261 agctggggtt tgtgaccctg cactctgaca aagccctcac cgaaaggtgg gggtttagtc
     4321 atttggccta gtaaagcaga acagaggcga tttgtcacgc aaaggcccgc cgcccgaggg
     4381 gaaacaggcg gcgggccaat ggctcggcgt tgttagctgg gggtcgcacc agggaagcac
     4441 agaagccctt ccctgagcag ggtttcaccg tgtggtgtcc cttcgtcctt gaaggcgaac
     4501 tggtaggcca ggccgtaatg ctcctgcacc atcttttgct catccaccgc gacaaagaaa
     4561 cggcgggtgt tcgccgggct gctgagggtc tgccactcgc cggacgcgcc caccgcatag
     4621 gcgtctccgg ctccacattc ctcggcctgc tgatactgat acttcgtcca cagcatcacc
     4681 gccttatcgg gcacctgcgc gaagctcacc agcgccccga accgcccggc gctcgtggtg
     4741 aaggtggacg actgcaccat gtcggacagt tccagcgtgg caggcgcggg cgtgttcgtc
     4801 ggcgcgtaca ccgtccccgc ctcgcacacg accagcgggg cggcccccgt gccgaactcg
     4861 gcggcgctgc cgtttgccag cgtgagcgcc gaagacgcca ccgcgcaggt acccatgggt
     4921 gccgaccacc agcaggtcca cgccgtgcgc cgcctcggtg agcacctcga ttggacggcc
     4981 cctcattagg ggcaaggacg tccggcatgc taaactcgcc gcgcgcttcc ccggaagcgc
     5041 gcacaccgcc tccgtcatcc ggcgggtgga acgcttcttc gaccgtcatc ctcttcggcc
     5101 agctgatgtc gcccgggtcg ttctgacgct ccttcccgcc gcgcagccac gcgaatttat
     5161 ccttgaccgg accaactgga agtatgggca gacggacgtg aacgtcttgt tgctggccgt
     5221 catttggcgg gacgtcgcca tccccctgct ctacgagttg ctgccccatg ggggcagcag
     5281 cgacaccgag attcggcaca ccctgatgga cgatgccctg tgcctgctgt ccgccgctga
     5341 catccgggtg ctgtattagc cgaccgcgaa ttcccagctt gagtaggaca aatccgccga
     5401 gcttcgacga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     5461 accaccgttg atatatccca atcgcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     5521 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     5581 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     5641 gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     5701 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     5761 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     5821 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     5881 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     5941 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     6001 attcaggttc atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct taatgaatta
     6061 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     6121 ccttaaacgc ctggtgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt
     6181 cgtcgaggcg gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct
     6241 tgcggagaac tgtgaatgca tcccgcaaga ggcccggcag t
//
