LOCUS       sequence-92571-e        5746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains BioBrick prefixed and suffixed insert with a modified lac 
            promoter, Shine-Delgarno sequence, 5'.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  TITLE     BioBrick(TM) compatible plasmids for Rhodococcus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5746)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSRKBB-empty
FEATURES             Location/Qualifiers
     source          1..5746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3488..4282
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3542..3561)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4152..4171
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4566..5154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5055..5074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5308..5325
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5476..5497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5512..5517,5518..5535,5536..5542)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      5645..5654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5664..5681
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
       61 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
      121 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
      181 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
      241 tttcttgccg ccaaggatct gatggcgcag gggatcaaga tcagtactgc ggcgtcgctg
      301 atcgccctgg cgacgttgtg cgggtggctt gtccctgagg gcgctgcgac agatagctaa
      361 aaatctgggt caggatcgcc gtagagcgcg cgtcgtcgat tggaggcttc ccctttggtt
      421 gacggtcttc aatcgctcta cggcgatcct gacgcttttt tgttgcgtac cgtcgatcgt
      481 tttatttctg tcgatcccga aaaagttttt gccttttgta aaaaacttct cggtcgcccc
      541 gcaaattttc gattccagat tttttaaaaa ccaagccaga aatacgacac accgtttgca
      601 gataatctgt ctttcggaaa aatcaagtgc gatacaaaat ttttagcacc cctgagctgc
      661 gcaaagtccc gcttcgtgaa aattttcgtg ccgcgtgatt ttccgccaaa aactttaacg
      721 aacgttcgtt ataatggtgt catgaccttc acgacgaagt actaaaattg gcccgaatca
      781 tcagctatgg atctctctga tgtcgcgctg gagtccgacg cgctcgatgc tgccgtcgat
      841 ttaaaaacgg tgatcggatt tttccgagct ctcgatacga cggacgcgcc agcatcacga
      901 gactgggcca gtgccgcgag cgacctagaa actctcgtgg cggatcttga ggagctggct
      961 gacgagctgc gtgctcggcc agcgccagga ggacgcacag tagtggagga tgcaatcagt
     1021 tgcgcctact gcggtggcct gattcctccc cggcctgacc cgcgaggacg gcgcgcaaaa
     1081 tattgctcag atgcgtgtcg tgccgcagcc agccgcgagc gcgccaacaa acgccacgcc
     1141 gaggagctgg aggcggctag gtcgcaaatg gcgctggaag tgcgtccccc gagcgaaatt
     1201 ttggcaatgg tcgtcacaga gctggaagcg gcagcgagaa ttatcgcgat cgtggcggtg
     1261 cccgcaggca tgacaaacat cgtaaatgcc gcgtttcgtg tgccgtggcc gcccaggacg
     1321 tgtcagcgcc gccaccacct gcaccgaatc ggcagcagcg tcgcgcgtcg aaaaagcgca
     1381 caggcggcaa gaagcgataa gctgcacgaa tacctgaaaa atgttgaacg ccccgtgagc
     1441 ggtaactcac agggcgtcgg ctaaccccca gtccaaacct gggagaaagc gctcaaaaat
     1501 gactctagcg gattcacgag acattgacac accggcctgg aaattttccg ctgatctgtt
     1561 cgacacccat cccgagctcg cgctgcgatc acgtggctgg acgagcgaag accgccgcga
     1621 gttcctcgct cacctgggca gagaaaattt ccagggcagc aagacccgcg acttcgccag
     1681 cgcttggatc aaagacccgg acacggagaa acacagccga agttataccg agttggttca
     1741 aaatcgcttg cccggtgcca gtatgttgct ctgacgcacg cgcagcacgc agccgtgctt
     1801 gtcctggaca ttgatgtgcc gagccaccag gccggcggga aaatcgagca cgtaaacccc
     1861 gaggtctacg cgattttgga gcgctgggca cgcctggaaa aagcgccagc ttggatcggc
     1921 gtgaatccac tgagcgggaa atgccagctc atctggctca ttgatccggt gtatgccgca
     1981 gcaggcatga gcagcccgaa tatgcgcctg ctggctgcaa cgaccgagga aatgacccgc
     2041 gttttcggcg ctgaccaggc tttttcacat aggctgagcc gtggccactg cactctccga
     2101 cgatcccagc cgtaccgctg gcatgcccag cacaatcgcg tggatcgcct agctgatctt
     2161 atggaggttg ctcgcatgat ctcaggcaca gaaaaaccta aaaaacgcta tgagcaggag
     2221 ttttctagcg gacgggcacg tatcgaagcg gcaagaaaag ccactgcgga agcaaaagca
     2281 cttgccacgc ttgaagcaag cctgccgagc gccgctgaag cgtctggaga gctgatcgac
     2341 ggcgtccgtg tcctctggac tgctccaggg cgtgccgccc gtgatgagac ggcttttcgc
     2401 cacgctttga ctgtgggata ccagttaaaa gcggctggtg agcgcctaaa agacaccaag
     2461 ggtcatcgag cctacgagcg tgcctacacc gtcgctcagg cggtcggagg aggccgtgag
     2521 cctgatctgc cgccggactg tgaccgccag acggattggc cgcgacgtgt gcgcggctac
     2581 gtcgctaaag gccagccagt cgtccctgct cgtcagacag agacgcagag ccagccgagg
     2641 cgaaaagctc tggccactat gggaagacgt ggcggtaaaa aggccgcaga acgctggaaa
     2701 gacccaaaca gtgagtacgc ccgagcacag cgagaaaaac tagctaagtc cagtcaacga
     2761 caagctagga aagctaaagg aaatcgcttg accattgcag gttggtttat gactgttgag
     2821 ggagagactg gctcgtggcc gacaatcaat gaagctatgt ctgaatttag cgtgtcacgt
     2881 cagaccgtga atagagcact taaggtctgc gggcattgaa cttccacgag gacgccgaaa
     2941 gcttcccagt aaatgtgcca tctcgtaggc agaaaacggt tcccccgtag ggtctctctc
     3001 ttggcctcct ttctaggtcg ggctgattgc tcttgaagct ctctaggggg gctcacacca
     3061 taggcagata acgttcccca ccggctcgcc tcgtaagcgc acaaggactg ctcccaaaga
     3121 gcttcaaagc cactgccgcg actgccttcg cgaagccttg ccccgcggaa atttcctcca
     3181 ccgagttcgt gcacacccct atgccaagct tctttcaccc taaattcgag agattggatt
     3241 cttaccgtgg aaattcttcg caaaaatcgt cccctgatcg cccttgcgac gttggcgtcg
     3301 gtgccgctgg ttgcgcttgg cttgaccgac ttgatctgat cctcattcgc cgtggcagcg
     3361 tgggtcgaat gagaatacga atggattggt cagggatttt ttcccgaagg gcactaattt
     3421 tgctaaagta agtgacgaag aagttcagcg ggcacaggat caagagacag gatgaggatc
     3481 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3541 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3601 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3661 tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3721 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3781 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3841 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3901 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3961 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat
     4021 gcccgacggc gaggatctcg tcgtgaccca cggcgatgcc tgcttgccga atatcatggt
     4081 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4141 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4201 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4261 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4321 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4381 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4441 ttcttcgccc accccaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4501 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4561 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4621 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4681 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4741 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4801 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4861 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4921 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     4981 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5041 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5101 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5161 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5221 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5281 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5341 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5401 aggtttcccg actggagaat tcatcctgaa cttatctaga cccgactgga aagcgggcag
     5461 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5521 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgtc tagtagaagg aggagatctg
     5581 gatccatatg ggcggttgat aagatatcga aaagcttgat ggtaccgagg tcgacgacat
     5641 gcatccatgg cggccgcctc gagcatcacc atcaccatca ctgattctcc ttacgcatct
     5701 gtgcggtatt tcacaccgca tgtactagtt aacttcagga cctgca
//
