LOCUS       sequence-444744-        5740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with mCherry integrated in the multiple 
            cloning site (SmR). There is a SmaI immediately upstream of mCherry 
            start codon, which allows for C-terminal mCherry fusions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5740)
  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 5740)
  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: pHBJT034
FEATURES             Location/Qualifiers
     source          1..5740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(792..809)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(792..808)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(801..823)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1017..1036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1136..1158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1196..1214)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1282..1384
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3373..3595
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3643..4593
                     /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     5492..5509
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5626..5647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5662..5667,5668..5685,5686..5692)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5700..5716
                     /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     5705..5727
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5724..5740
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5724..5740
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggatggtat
       61 caaaaggaga ggaagataat atggcgatca tcaaagagtt catgcgtttt aaagtgcaca
      121 tggaaggctc cgtgaacggt cacgagttcg aaattgaggg cgagggcgag ggccgtccgt
      181 atgaaggcac gcaaactgct aaactcaagg tcacgaaagg cggaccactg ccttttgcct
      241 gggacatcct gtctccgcag tttatgtacg gcagcaaagc gtacgttaaa catccggcag
      301 atattccgga ctacctgaag ttgtccttcc cggagggttt taagtgggag cgtgttatga
      361 atttcgagga cggcggtgtc gtgaccgtta cccaagacag cagcctgcaa gatggcgagt
      421 tcatctataa agttaagctg cgtggtacga acttcccgtc tgatggcccg gtgatgcaga
      481 aaaagaccat gggttgggaa gcgagcagcg agcgcatgta cccggaagac ggtgcgctga
      541 agggtgagat caaacagcgc ctgaagttga aggacggtgg ccactacgat gcagaagtaa
      601 aaaccaccta taaggctaaa aaaccggttc agttgccggg tgcctataac gtgaatatta
      661 agttagacat caccagccat aacgaagact acaccattgt ggaacaatat gagcgcgcgg
      721 agggtcgtca cagcaccggt ggtatggatg aactgtacaa gtaatgataa tagggtaccg
      781 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      841 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      901 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
      961 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     1021 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     1081 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1141 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     1201 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1261 ggcacttttc ggggaaatgt gtgatcggca cgtaagaggt tccaactttc accataatga
     1321 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     1381 taaattgaat cgaactaata ttttttttgg tgaatcgcat tctgactggt tgcctgtcag
     1441 aggcggagaa tctggtgatt ttgtttttcg acgtggtgac gggcatgcct tcgcgaaaat
     1501 cgcacctgct tcccgccgcg gtgagctcgc tggagagcgt gaccgcctca tttggctcaa
     1561 aggtcgaggt gtggcttgcc ccgaggtcat caactggcag gaggaacagg agggtgcatg
     1621 cttggtgata acggcaattc cgggagtacc ggcggctgat ctgtctggag cggatttgct
     1681 caaagcgtgg ccgtcaatgg ggcagcaact tggcgctgtt cacagcctat cggttgatca
     1741 atgtccgttt gagcgcaggc tgtcgcgaat gttcggacgc gccgttgatg tggtgtcccg
     1801 caatgccgtc aatcccgact tcttaccgga cgaggacaag agtacgccgc agctcgatct
     1861 tttggctcgt gtcgaacgag agctaccggt gcggctcgac caagagcgca ccgatatggt
     1921 tgtttgccat ggtgatccct gcatgccgaa cttcatggtg gaccctaaaa ctcttcaatg
     1981 cacgggtctg atcgaccttg ggcggctcgg aacagcagat cgctatgccg atttggcact
     2041 catgattgct aacgccgaag agaactgggc agcgccagat gaagcagagc gcgccttcgc
     2101 tgtcctattc aatgtattgg ggatcgaagc ccccgaccgc gaacgccttg ccttctatct
     2161 gcgattggac cctctgactt ggggttgatg ttcatgccgc ctgtttttcc tgctcattgg
     2221 cacgtttcgc aacctgttct cattgcggac accttttcca gcctcgtttg gaaagtttca
     2281 ttgccagacg ggactcctgc aatcgtcaag ggattgaaac ctatagaaga cattgctgat
     2341 gaactgcgcg gggccgacta tctggtatgg cgcaatggga ggggagcagt ccggttgctc
     2401 ggtcgtgaga acaatctgat gttgctcgaa tatgccgggg agcgaatgct ctctcacatc
     2461 gttgccgagc acggcgacta ccaggcgacc gaaattgcag cggaactaat ggcgaagctg
     2521 tatgccgcat ctgaggaacc cctgccttct gcccttctcc cgatccggga tcgctttgca
     2581 gctttgtttc agcgggcgcg cgatgatcaa aacgcaggtt gtcaaactga ctacgtccac
     2641 gcggcgatta tagccgatca aatgatgagc aatgcctcgg aactgcgtgg gctacatggc
     2701 gatctgcatc atgaaaacat catgttctcc agtcgcggct ggctggtgat agatcccgtc
     2761 ggtctggtcg gtgaagtggg ctttggcgcc gccaatatgt tctacgatcc ggctgacaga
     2821 gacgaccttt gtctcgatcc tagacgcatt gcacagatgg cggacgcatt ctctcgtgcg
     2881 ctggacgtcg atccgcgtcg cctgctcgac caggcgtacg cttatgggtg cctttccgca
     2941 gcttggaacg cggatggaga agaggagcaa cgcgatctag ctatcgcggc cgcgatcaag
     3001 caggtgcgac agacgtcata ctagcctcgc taacggattc accactccgc agacccgcca
     3061 taaaacgccc tgagaagccc gtgacgggct tttcttgtat tatgggtagt ttccttgcat
     3121 gaatccataa aaggcgcctg tagtgccatt tacccccatt cactgccaga gccgtgagcg
     3181 cagcgaactg aatgtcacga aaaagacagc gactcaggtg cctgatggtc ggagacaaaa
     3241 ggaatattca gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa
     3301 ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact
     3361 gactaaagta gtgagttata cacagggctg ggatctattc tttttatctt tttttattct
     3421 ttctttattc tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta
     3481 gcggaattta cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat
     3541 ttacagatac ccacaactca aaggaaaagg actagtaatt atcattgact agcccatctc
     3601 aattggtata gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt
     3661 ttcaaagcaa atgaactagc gattagtcgc tatgacttaa cggagcatga aaccaagcta
     3721 attttatgct gtgtggcact actcaacccc acgattgaaa accctacaag gaaagaacgg
     3781 acggtatcgt tcacttataa ccaatacgct cagatgatga acatcagtag ggaaaatgct
     3841 tatggtgtat tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcaggaat
     3901 cctttggtta aaggctttga gattttccag tggacaaact atgccaagtt ctcaagcgaa
     3961 aaattagaat tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc
     4021 ataaaatata atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat
     4081 gagtggttat taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc
     4141 cttgatgaat ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt
     4201 aaccaatggg ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg
     4261 gtggttgata agcgaggccg cccgactgat acgttgattt tccaagttga actagataga
     4321 caaatggatc tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata
     4381 ccaacaacca ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat
     4441 gctttaactg caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa
     4501 agtaagtatg atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta
     4561 gagaacatac tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag
     4621 tgaagcatca agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg
     4681 aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag atagatacat cagagctttt
     4741 acgagttttt ggtgcattca aagctgttca ccatgaacag atcgacaatg taacagatga
     4801 acagcatgta acacctaata gaacaggtga aaccagtaaa acaaagcaac tagaacatga
     4861 aattgaacac ctgagacaac ttgttacagc tcaacagtca cacatagaca gcctgaaaca
     4921 ggcgatgctg cttatcgaat caaagctgcc gacaacacgg gagccagtga cgcctcccgt
     4981 ggggaaaaaa tcatggcaat tctggaagaa atagcgcttt cagccggcaa accggctgaa
     5041 gccggatctg cgattctgat aacaaactag caacaccaga acagcccgtt tgcgggcagc
     5101 aaaacccgta cttttggacg ttccggcggt tttttgtggc gagtggtgtt cgggcggtgc
     5161 gcgcaagatc cattatgtta aacgggcgag tttacatctc aaaaccgccc gcttaacacc
     5221 atcagaaatc ctcagcgcga ttttaagcac caaccccccc cccccccccc cccccgtaac
     5281 acccaaatcc atactgaaag tggctttgtt gaataaatca gatttcgggt aagtctccaa
     5341 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5401 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5461 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5521 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5581 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     5641 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5701 tgtgagcgga taacaatttc acacaggaaa cagctatgac
//
