LOCUS       sequence-444700-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with same multiple cloning site as pUC19 and
            pSU19 (SmR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5008)
  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 5008)
  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: pHBJT023
FEATURES             Location/Qualifiers
     source          1..5008
                     /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     2024..2041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2158..2179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2194..2199,2200..2217,2218..2224)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2232..2248
                     /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     2237..2259
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2256..2272
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2256..2272
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2268..2591
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2285..2341
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2342..2359)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2342..2358)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2351..2373)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2567..2586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2686..2708
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2746..2764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2832..2934
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      join(4923..5008,1..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 cccccccgta acacccaaat ccatactgaa agtggctttg ttgaataaat cagatttcgg
     1861 gtaagtctcc aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1921 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1981 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2041 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2101 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2161 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2221 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2281 cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg agctcgaatt
     2341 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2401 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2461 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2521 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     2581 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2641 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2701 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     2761 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2821 ggggaaatgt gtgatcggca cgtaagaggt tccaactttc accataatga aataagatca
     2881 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaattgaat
     2941 cgaactaata ttttttttgg tgaatcgcat tctgactggt tgcctgtcag aggcggagaa
     3001 tctggtgatt ttgtttttcg acgtggtgac gggcatgcct tcgcgaaaat cgcacctgct
     3061 tcccgccgcg gtgagctcgc tggagagcgt gaccgcctca tttggctcaa aggtcgaggt
     3121 gtggcttgcc ccgaggtcat caactggcag gaggaacagg agggtgcatg cttggtgata
     3181 acggcaattc cgggagtacc ggcggctgat ctgtctggag cggatttgct caaagcgtgg
     3241 ccgtcaatgg ggcagcaact tggcgctgtt cacagcctat cggttgatca atgtccgttt
     3301 gagcgcaggc tgtcgcgaat gttcggacgc gccgttgatg tggtgtcccg caatgccgtc
     3361 aatcccgact tcttaccgga cgaggacaag agtacgccgc agctcgatct tttggctcgt
     3421 gtcgaacgag agctaccggt gcggctcgac caagagcgca ccgatatggt tgtttgccat
     3481 ggtgatccct gcatgccgaa cttcatggtg gaccctaaaa ctcttcaatg cacgggtctg
     3541 atcgaccttg ggcggctcgg aacagcagat cgctatgccg atttggcact catgattgct
     3601 aacgccgaag agaactgggc agcgccagat gaagcagagc gcgccttcgc tgtcctattc
     3661 aatgtattgg ggatcgaagc ccccgaccgc gaacgccttg ccttctatct gcgattggac
     3721 cctctgactt ggggttgatg ttcatgccgc ctgtttttcc tgctcattgg cacgtttcgc
     3781 aacctgttct cattgcggac accttttcca gcctcgtttg gaaagtttca ttgccagacg
     3841 ggactcctgc aatcgtcaag ggattgaaac ctatagaaga cattgctgat gaactgcgcg
     3901 gggccgacta tctggtatgg cgcaatggga ggggagcagt ccggttgctc ggtcgtgaga
     3961 acaatctgat gttgctcgaa tatgccgggg agcgaatgct ctctcacatc gttgccgagc
     4021 acggcgacta ccaggcgacc gaaattgcag cggaactaat ggcgaagctg tatgccgcat
     4081 ctgaggaacc cctgccttct gcccttctcc cgatccggga tcgctttgca gctttgtttc
     4141 agcgggcgcg cgatgatcaa aacgcaggtt gtcaaactga ctacgtccac gcggcgatta
     4201 tagccgatca aatgatgagc aatgcctcgg aactgcgtgg gctacatggc gatctgcatc
     4261 atgaaaacat catgttctcc agtcgcggct ggctggtgat agatcccgtc ggtctggtcg
     4321 gtgaagtggg ctttggcgcc gccaatatgt tctacgatcc ggctgacaga gacgaccttt
     4381 gtctcgatcc tagacgcatt gcacagatgg cggacgcatt ctctcgtgcg ctggacgtcg
     4441 atccgcgtcg cctgctcgac caggcgtacg cttatgggtg cctttccgca gcttggaacg
     4501 cggatggaga agaggagcaa cgcgatctag ctatcgcggc cgcgatcaag caggtgcgac
     4561 agacgtcata ctagcctcgc taacggattc accactccgc agacccgcca taaaacgccc
     4621 tgagaagccc gtgacgggct tttcttgtat tatgggtagt ttccttgcat gaatccataa
     4681 aaggcgcctg tagtgccatt tacccccatt cactgccaga gccgtgagcg cagcgaactg
     4741 aatgtcacga aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca
     4801 gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt
     4861 gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta
     4921 gtgagttata cacagggctg ggatctattc tttttatctt tttttattct ttctttattc
     4981 tataaattat aaccacttga atataaac
//
