LOCUS       sequence-444735-        5004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with same multiple cloning site as pUC18 and
            pSU18 (SmR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5004)
  AUTHORS   Braun HG, Kanwal N, Rivera Lopez LF, Thomassin JL
  TITLE     Generation of a plasmid series for rapid sub-cloning and use in 
            various Enterobacteriaceae.
  JOURNAL   J Biosci Bioeng. 2024 Sep 6:S1389-1723(24)00253-6. doi: 
            10.1016/j.jbiosc.2024.08.006.
  PUBMED    39244484
REFERENCE   2  (bases 1 to 5004)
  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: "J Biosci 
            Bioeng. 2024 Sep 6:S1389-1723(24)00253-6. doi: 
            10.1016/j.jbiosc.2024.08.006."
COMMENT     Sequence Label: pHBJT024
FEATURES             Location/Qualifiers
     source          1..5004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             185..1135
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     2021..2038
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2155..2176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2191..2196,2197..2214,2215..2221)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2229..2245
                     /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     2234..2256
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2253..2269
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2253..2269
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2270..2587
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDA
                     A"
     misc_feature    2278..2334
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2338..2355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2338..2354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2347..2369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2563..2582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2682..2704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2742..2760)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2828..2930
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      join(4919..5004,1..137)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt ctagcagaat ttacaagttt
       61 tccagcaaag gtctagcaga atttacagat acccacaact caaaggaaaa ggactagtaa
      121 ttatcattga ctagcccatc tcaattggta tagtgattaa aatcacctag accaattgag
      181 atgtatgtct gaattagttg ttttcaaagc aaatgaacta gcgattagtc gctatgactt
      241 aacggagcat gaaaccaagc taattttatg ctgtgtggca ctactcaacc ccacgattga
      301 aaaccctaca aggaaagaac ggacggtatc gttcacttat aaccaatacg ctcagatgat
      361 gaacatcagt agggaaaatg cttatggtgt attagctaaa gcaaccagag agctgatgac
      421 gagaactgtg gaaatcagga atcctttggt taaaggcttt gagattttcc agtggacaaa
      481 ctatgccaag ttctcaagcg aaaaattaga attagttttt agtgaagaga tattgcctta
      541 tcttttccag ttaaaaaaat tcataaaata taatctggaa catgttaagt cttttgaaaa
      601 caaatactct atgaggattt atgagtggtt attaaaagaa ctaacacaaa agaaaactca
      661 caaggcaaat atagagatta gccttgatga atttaagttc atgttaatgc ttgaaaataa
      721 ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa ccaataagta aagatttaaa
      781 cacttacagc aatatgaaat tggtggttga taagcgaggc cgcccgactg atacgttgat
      841 tttccaagtt gaactagata gacaaatgga tctcgtaacc gaacttgaga acaaccagat
      901 aaaaatgaat ggtgacaaaa taccaacaac cattacatca gattcctacc tacataacgg
      961 actaagaaaa acactacacg atgctttaac tgcaaaaatt cagctcacca gttttgaggc
     1021 aaaatttttg agtgacatgc aaagtaagta tgatctcaat ggttcgttct catggctcac
     1081 gcaaaaacaa cgaaccacac tagagaacat actggctaaa tacggaagga tctgaggttc
     1141 ttatggctct tgtatctatc agtgaagcat caagactaac aaacaaaagt agaacaactg
     1201 ttcaccgtta catatcaaag ggaaaactgt ccatatgcac agatgaaaac ggtgtaaaaa
     1261 agatagatac atcagagctt ttacgagttt ttggtgcatt caaagctgtt caccatgaac
     1321 agatcgacaa tgtaacagat gaacagcatg taacacctaa tagaacaggt gaaaccagta
     1381 aaacaaagca actagaacat gaaattgaac acctgagaca acttgttaca gctcaacagt
     1441 cacacataga cagcctgaaa caggcgatgc tgcttatcga atcaaagctg ccgacaacac
     1501 gggagccagt gacgcctccc gtggggaaaa aatcatggca attctggaag aaatagcgct
     1561 ttcagccggc aaaccggctg aagccggatc tgcgattctg ataacaaact agcaacacca
     1621 gaacagcccg tttgcgggca gcaaaacccg tacttttgga cgttccggcg gttttttgtg
     1681 gcgagtggtg ttcgggcggt gcgcgcaaga tccattatgt taaacgggcg agtttacatc
     1741 tcaaaaccgc ccgcttaaca ccatcagaaa tcctcagcgc gattttaagc accaaccccc
     1801 ccccgtaaca cccaaatcca tactgaaagt ggctttgttg aataaatcag atttcgggta
     1861 agtctccaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1921 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1981 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2041 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2101 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2161 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2221 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     2281 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggcact
     2341 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2401 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2461 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     2521 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     2581 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2641 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2701 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     2761 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2821 aaatgtgtga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     2881 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa ttgaatcgaa
     2941 ctaatatttt ttttggtgaa tcgcattctg actggttgcc tgtcagaggc ggagaatctg
     3001 gtgattttgt ttttcgacgt ggtgacgggc atgccttcgc gaaaatcgca cctgcttccc
     3061 gccgcggtga gctcgctgga gagcgtgacc gcctcatttg gctcaaaggt cgaggtgtgg
     3121 cttgccccga ggtcatcaac tggcaggagg aacaggaggg tgcatgcttg gtgataacgg
     3181 caattccggg agtaccggcg gctgatctgt ctggagcgga tttgctcaaa gcgtggccgt
     3241 caatggggca gcaacttggc gctgttcaca gcctatcggt tgatcaatgt ccgtttgagc
     3301 gcaggctgtc gcgaatgttc ggacgcgccg ttgatgtggt gtcccgcaat gccgtcaatc
     3361 ccgacttctt accggacgag gacaagagta cgccgcagct cgatcttttg gctcgtgtcg
     3421 aacgagagct accggtgcgg ctcgaccaag agcgcaccga tatggttgtt tgccatggtg
     3481 atccctgcat gccgaacttc atggtggacc ctaaaactct tcaatgcacg ggtctgatcg
     3541 accttgggcg gctcggaaca gcagatcgct atgccgattt ggcactcatg attgctaacg
     3601 ccgaagagaa ctgggcagcg ccagatgaag cagagcgcgc cttcgctgtc ctattcaatg
     3661 tattggggat cgaagccccc gaccgcgaac gccttgcctt ctatctgcga ttggaccctc
     3721 tgacttgggg ttgatgttca tgccgcctgt ttttcctgct cattggcacg tttcgcaacc
     3781 tgttctcatt gcggacacct tttccagcct cgtttggaaa gtttcattgc cagacgggac
     3841 tcctgcaatc gtcaagggat tgaaacctat agaagacatt gctgatgaac tgcgcggggc
     3901 cgactatctg gtatggcgca atgggagggg agcagtccgg ttgctcggtc gtgagaacaa
     3961 tctgatgttg ctcgaatatg ccggggagcg aatgctctct cacatcgttg ccgagcacgg
     4021 cgactaccag gcgaccgaaa ttgcagcgga actaatggcg aagctgtatg ccgcatctga
     4081 ggaacccctg ccttctgccc ttctcccgat ccgggatcgc tttgcagctt tgtttcagcg
     4141 ggcgcgcgat gatcaaaacg caggttgtca aactgactac gtccacgcgg cgattatagc
     4201 cgatcaaatg atgagcaatg cctcggaact gcgtgggcta catggcgatc tgcatcatga
     4261 aaacatcatg ttctccagtc gcggctggct ggtgatagat cccgtcggtc tggtcggtga
     4321 agtgggcttt ggcgccgcca atatgttcta cgatccggct gacagagacg acctttgtct
     4381 cgatcctaga cgcattgcac agatggcgga cgcattctct cgtgcgctgg acgtcgatcc
     4441 gcgtcgcctg ctcgaccagg cgtacgctta tgggtgcctt tccgcagctt ggaacgcgga
     4501 tggagaagag gagcaacgcg atctagctat cgcggccgcg atcaagcagg tgcgacagac
     4561 gtcatactag cctcgctaac ggattcacca ctccgcagac ccgccataaa acgccctgag
     4621 aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat ccataaaagg
     4681 cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc gaactgaatg
     4741 tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa tattcagcga
     4801 tttgcccgag cttgcgaggg tgctacttaa gcctttaggg ttttaaggtc tgttttgtag
     4861 aggagcaaac agcgtttgcg acatcctttt gtaatactgc ggaactgact aaagtagtga
     4921 gttatacaca gggctgggat ctattctttt tatctttttt tattctttct ttattctata
     4981 aattataacc acttgaatat aaac
//
