LOCUS       sequence-385598-        7001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7001)
  AUTHORS   Smith K, Youngman P
  TITLE     Use of a new integrational vector to investigate 
            compartment-specific expression of the Bacillus subtilis spoIIM 
            gene.
  JOURNAL   Biochimie.   . 74(7-8):705-11.
  PUBMED    1391050
REFERENCE   2  (bases 1 to 7001)
  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: "Biochimie. 
             . 74(7-8):705-11."
COMMENT     Sequence Label: pKSV7
FEATURES             Location/Qualifiers
     source          1..7001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             116..433
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDA
                     A"
     misc_feature    124..180
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(184..201)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(184..200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(193..215)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(409..428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     528..550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(588..606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        674..778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(779..847,848..1639)
                     /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(997..1016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1810..2398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2299..2318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2552..2569
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattcg agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt
      181 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
      241 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
      301 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
      361 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
      421 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
      481 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      541 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
      601 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
      661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2461 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2521 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2581 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgatt
     2641 tcactttttg cattctacaa actgcataac tcatatgtaa atcgctcctt tttaggtggc
     2701 acaaatgtga ggcattttcg ctctttccgg cgaggctagt tacccttaag ttattggtat
     2761 gactggtttt aagcgcaaaa aaagttgctt tttcgtacct attaatgtat cgttagaaaa
     2821 ccgactgtaa aaagtacagt cggcattatc tcatattata aaagccagtc attaggccta
     2881 tctgacaatt cctgaataga gttcataaac aatcctgcat gataaccatc acaaacagaa
     2941 tgatgtacct gtaaagatag cggtaaatat attgaattac ctttattaat gaattttcct
     3001 gctgtaataa tgggtagaag gtaattacta ttattattga tatttaagtt aaacccagta
     3061 aatgaagtcc atggaataat agaaagagaa aaagcatttt caggtatagg tgttttggga
     3121 aacaatttcc ccgaaccatt atatttctct acatcagaaa ggtataaatc ataaaactct
     3181 ttgaagtcat tctttacagg agtccaaata ccagagaatg ttttagatac accatcaaaa
     3241 attgtataaa gtggctctaa cttatcccaa taacctaact ctccgtcgct attgtaacca
     3301 gttctaaaag ctgtatttga gtttatcacc cttgtcacta agaaaataaa tgcagggtaa
     3361 aatttatatc cttcttgttt tatgtttcgg tataaaacac taatatcaat ttctgtggtt
     3421 atactaaaag tcgtttgttg gttcaaataa tgattaaata tctcttttct cttccaattg
     3481 tctaaatcaa ttttattaaa gttcatttga tatgcctcct aaatttttat ctaaagtgaa
     3541 tttaggaggc ttacttgtct gctttcttca ttagaatcaa tcctttttta aaagtcaata
     3601 ttactgtaac ataaatatat attttaaaaa tatcccactt tatccaattt tcgtttgttg
     3661 aactaatggg tgctttagtt gaagaataaa agaccacatt aaaaaatgtg gtcttttgtg
     3721 tttttttaaa ggatttgagc gtagcgaaaa atccttttct ttcttatctt gataataagg
     3781 gtaactattg ccgggataga ctgtaacatt ctcacgcata aaatcccctt tcattttcta
     3841 atgtaaatct attaccttat tattaattca attcgctcat aattaatcct ttttcttatt
     3901 acgcaaaatg gcccgattta agcacaccct ttattccgtt aatgcgccat gacagccatg
     3961 ataattacta atactaggag aagttaataa atacgtaacc aacatgatta acaattatta
     4021 gaggtcatcg ttcaaaatgg tatgcgtttt gacacatcca ctatatatcc gtgtcgttct
     4081 gtccactcct gaatcccatt ccagaaattc tctagcgatt ccagaagttt ctcagagtcg
     4141 gaaagttgac cagacattac gaactggcac agatggtcat aacctgaagg aagatcacga
     4201 taatttccaa gttttaaaca aggatatatt gcagtttaaa tttcctaaaa accaatccta
     4261 taaaatatat ggtaatatac cttataacat aagtacggat ataatacgca aaattgtttt
     4321 tgatagtata gctaatgaga tttatttaat cgtggaatac gggtttgcta aaagattatt
     4381 aaatacaaaa cgctcattgg cattactttt aatggcagaa gttgatattt ctatattaag
     4441 tatggttcca agagaatatt ttcatcctaa acctaaagtg aatagctcac ttatcagatt
     4501 aagtagaaaa aaatcaagaa tatcacacaa agataaacaa aagtataatt atttcgttat
     4561 gaaatgggtt aacaaagaat acaagaaaat atttacaaaa aatcaattta acaattcctt
     4621 aaaacatgca ggaattgacg atttaaacaa tattagcttt gaacaattct tatctctttt
     4681 caatagctat aaattattta ataagtaagt taagggatgc ataaactgca tcccttaact
     4741 tgtttttcgt gtgcctattt tttgtgaatc gattatgtct tttgcgcagt cggcttaaac
     4801 cagttttcgt tggtgcgaaa aaagagtgtc ttgtgacact cttaaattca aaatctatcg
     4861 gtcagattta taccgatttg attttatata ttcttgaata acatacgccg agttatcaca
     4921 taaaagcggg aaccaatcat caaatttaaa cttcattgca taatccatta aactcttaaa
     4981 ttctacgatt ccttgttcat caataaactc aatcatttct ttaattaatt tatatctatc
     5041 tgttgttgtt ttctttaata attcatcaac atctacaccg ccataaacta tcatatcttc
     5101 tttttgatat ttaaatttat taggatcgtc catgtgaagc atatatctca caagaccttt
     5161 cacacttcct gcaatctgcg gaatagtcgc attcaattct tctgtaatta tttttatctg
     5221 ttcataagat ttattaccct catacatcac tagaatatga taatgctctt ttttcatcct
     5281 atcttctgta tcagtatccc tatcatgtaa tggagacact acaaattgaa tgtgtaactc
     5341 ttttaaatac tctaaccact cggcttttgc tgattctgga tataaaacaa atgtccaatt
     5401 acgtcctctt gaatttttct tgttttcagt ttcttttatt acattttcgc tcatgatata
     5461 ataacggtgc taatacattt aacaaaattt agtcatagat aggcagcatg ccagtgctgt
     5521 ctatcttttt ttgtttaaaa tgcaccgtat tcctcctttg catatttttt tattagaata
     5581 ccggttgcat ctgatttgct aatattatat ttttctttga ttctatttaa tatctcattt
     5641 tcttctgttg taagtcttaa agtaacagca acttttttct cttcttttct atctacaacc
     5701 atcactgtac ctcccaacat ctgttttttt cactttaaca taaaaaacaa ccttttaaca
     5761 ttaaaaaccc aatatttatt tatttgtttg gacaatggac aatggacacc taggggggag
     5821 gtcgtagtac ccccctatgt tttctcccct aaataacccc aaaaatctaa gaaaaaaaga
     5881 cctcaaaaag gtctttaatt aacatctcaa atttcgcatt tattccaatt tcctttttgc
     5941 gtgtgatgcg ctgcgtccat taaaaatcct agagctttgc aaccgaaagt taatagctgt
     6001 cgctactact ttcgcttacg ctctaagtat attttaagga ctgtcacacg caaaaagttt
     6061 tctcggcata aaagtacctc tacatctcta aatcgtctgt acgctgtttc tcacgctttc
     6121 tatcgacctt ctggacatta tcctgtacaa catccataaa ctgtcccaca cgctcgaatt
     6181 tggaatcatt aaagaatttc tctttaagcc tattaaaccc tttctcaaac ccagggaaat
     6241 tcgccctcgc agcacgatat aaagtcactg tactatcttg aaatttctct gatacattca
     6301 actgctcatt caaactatca ttctctcgct ttaatttatt aacctcttta cttttttcgt
     6361 gatacccctc tttccatgta ttcactactt ctttcaaact ctctctacgt ttttttaatt
     6421 cttgattttc tgtgtaatag tctgtgctct taatattttc gtaatcatca acaatccgtt
     6481 cgaagagatt gtttcttgca ggcgttcaaa ttcatcagca gttaatatct ttctaccagt
     6541 ctcttcacgt ccagagaaca aacctgtacg ctcattttca taatcaaagg gtttcgtaga
     6601 cctcatatgc tctattccac tctgtaactg cttatttgcc ttctgtaact catccttaac
     6661 ttcttgcagt tcctgtttat gaaatacagt atctttcttg tactgatcca gttgctcaaa
     6721 aaaatctcgg tcagatgtta ctagcaactc atttacaaga acagcatctt tcctcgtttt
     6781 tcttgtacct gttttttgtg attcaataat ttctttgaca cgttcgttgt aatcaatatt
     6841 tttatcattt ttcaaatcat aattttcacg tgttcgctca tggtcaatat catcattcgt
     6901 tctacttttt cgctctcttt gattatgaaa ttgcatgcct tttagtccag ctggcacgac
     6961 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa t
//
