LOCUS       sequence-444689-        4623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with mCherry integrated in the multiple 
            cloning site (GmR). 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 4623)
  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 4623)
  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: pHBJT033
FEATURES             Location/Qualifiers
     source          1..4623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      387..610
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             658..1608
                     /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     2495..2512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2629..2650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2665..2670,2671..2688,2689..2695)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2703..2719
                     /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     2708..2730
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2727..2743
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2727..2743
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3535..3552)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3535..3551)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3544..3566)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3760..3779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3879..3901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3939..3957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4025..4127
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4128..4623,1..38)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 tgatgcactt tgatatcgac ccaagtaccg ccacctaacc tcgctaacgg attcaccact
       61 ccgcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt gtattatggg
      121 tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc cattcactgc
      181 cagagccgtg agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat
      241 ggtcggagac aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc
      301 ctttagggtt ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt
      361 aatactgcgg aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta
      421 tcttttttta ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa
      481 acacacaaag gtctagcgga atttacagag ggtctagcag aatttacaag ttttccagca
      541 aaggtctagc agaatttaca gatacccaca actcaaagga aaaggactag taattatcat
      601 tgactagccc atctcaattg gtatagtgat taaaatcacc tagaccaatt gagatgtatg
      661 tctgaattag ttgttttcaa agcaaatgaa ctagcgatta gtcgctatga cttaacggag
      721 catgaaacca agctaatttt atgctgtgtg gcactactca accccacgat tgaaaaccct
      781 acaaggaaag aacggacggt atcgttcact tataaccaat acgctcagat gatgaacatc
      841 agtagggaaa atgcttatgg tgtattagct aaagcaacca gagagctgat gacgagaact
      901 gtggaaatca ggaatccttt ggttaaaggc tttgagattt tccagtggac aaactatgcc
      961 aagttctcaa gcgaaaaatt agaattagtt tttagtgaag agatattgcc ttatcttttc
     1021 cagttaaaaa aattcataaa atataatctg gaacatgtta agtcttttga aaacaaatac
     1081 tctatgagga tttatgagtg gttattaaaa gaactaacac aaaagaaaac tcacaaggca
     1141 aatatagaga ttagccttga tgaatttaag ttcatgttaa tgcttgaaaa taactaccat
     1201 gagtttaaaa ggcttaacca atgggttttg aaaccaataa gtaaagattt aaacacttac
     1261 agcaatatga aattggtggt tgataagcga ggccgcccga ctgatacgtt gattttccaa
     1321 gttgaactag atagacaaat ggatctcgta accgaacttg agaacaacca gataaaaatg
     1381 aatggtgaca aaataccaac aaccattaca tcagattcct acctacataa cggactaaga
     1441 aaaacactac acgatgcttt aactgcaaaa attcagctca ccagttttga ggcaaaattt
     1501 ttgagtgaca tgcaaagtaa gtatgatctc aatggttcgt tctcatggct cacgcaaaaa
     1561 caacgaacca cactagagaa catactggct aaatacggaa ggatctgagg ttcttatggc
     1621 tcttgtatct atcagtgaag catcaagact aacaaacaaa agtagaacaa ctgttcaccg
     1681 ttacatatca aagggaaaac tgtccatatg cacagatgaa aacggtgtaa aaaagataga
     1741 tacatcagag cttttacgag tttttggtgc attcaaagct gttcaccatg aacagatcga
     1801 caatgtaaca gatgaacagc atgtaacacc taatagaaca ggtgaaacca gtaaaacaaa
     1861 gcaactagaa catgaaattg aacacctgag acaacttgtt acagctcaac agtcacacat
     1921 agacagcctg aaacaggcga tgctgcttat cgaatcaaag ctgccgacaa cacgggagcc
     1981 agtgacgcct cccgtgggga aaaaatcatg gcaattctgg aagaaatagc gctttcagcc
     2041 ggcaaaccgg ctgaagccgg atctgcgatt ctgataacaa actagcaaca ccagaacagc
     2101 ccgtttgcgg gcagcaaaac ccgtactttt ggacgttccg gcggtttttt gtggcgagtg
     2161 gtgttcgggc ggtgcgcgca agatccatta tgttaaacgg gcgagtttac atctcaaaac
     2221 cgcccgctta acaccatcag aaatcctcag cgcgatttta agcaccaacc ccccccccgt
     2281 aacacccaaa tccatactga aagtggcttt gttgaataaa tcagatttcg ggtaagtctc
     2341 caacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2461 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2521 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2581 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     2641 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     2701 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     2761 tgcatgcctg caggtcgact ctagaggatc cccgggatgg tatcaaaagg agaggaagat
     2821 aatatggcga tcatcaaaga gttcatgcgt tttaaagtgc acatggaagg ctccgtgaac
     2881 ggtcacgagt tcgaaattga gggcgagggc gagggccgtc cgtatgaagg cacgcaaact
     2941 gctaaactca aggtcacgaa aggcggacca ctgccttttg cctgggacat cctgtctccg
     3001 cagtttatgt acggcagcaa agcgtacgtt aaacatccgg cagatattcc ggactacctg
     3061 aagttgtcct tcccggaggg ttttaagtgg gagcgtgtta tgaatttcga ggacggcggt
     3121 gtcgtgaccg ttacccaaga cagcagcctg caagatggcg agttcatcta taaagttaag
     3181 ctgcgtggta cgaacttccc gtctgatggc ccggtgatgc agaaaaagac catgggttgg
     3241 gaagcgagca gcgagcgcat gtacccggaa gacggtgcgc tgaagggtga gatcaaacag
     3301 cgcctgaagt tgaaggacgg tggccactac gatgcagaag taaaaaccac ctataaggct
     3361 aaaaaaccgg ttcagttgcc gggtgcctat aacgtgaata ttaagttaga catcaccagc
     3421 cataacgaag actacaccat tgtggaacaa tatgagcgcg cggagggtcg tcacagcacc
     3481 ggtggtatgg atgaactgta caagtaatga taatagggta ccgagctcga attcactggc
     3541 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     3601 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3661 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     3721 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3781 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3901 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3961 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4021 tgtgtgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     4081 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg ttacgcagca
     4141 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg
     4201 gcatcattcg cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg
     4261 atcttttcgg tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg
     4321 attacctcgg gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag
     4381 aagcggttgt tggcgctctc gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg
     4441 agatctatat ctatgatctc gcagtctccg gcgagcaccg gaggcagggc attgccaccg
     4501 cgctcatcaa tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc
     4561 aagcagatta cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag
     4621 aag
//
