LOCUS       sequence-444686-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with mCherry integrated in the multiple 
            cloning site (Chlr). 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 4749)
  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 4749)
  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: pHBJT030
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(793..810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(793..809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..824)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1018..1037)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1137..1159
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1197..1215)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1283..1385
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1386..2045
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1452..1471)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2394..2616
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2664..3614
                     /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     4502..4519
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4636..4657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4672..4677,4678..4695,4696..4702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4710..4726
                     /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     4715..4737
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4734..4749,1)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4734..4749,1)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc cgggatggta
       61 tcaaaaggag aggaagataa tatggcgatc atcaaagagt tcatgcgttt taaagtgcac
      121 atggaaggct ccgtgaacgg tcacgagttc gaaattgagg gcgagggcga gggccgtccg
      181 tatgaaggca cgcaaactgc taaactcaag gtcacgaaag gcggaccact gccttttgcc
      241 tgggacatcc tgtctccgca gtttatgtac ggcagcaaag cgtacgttaa acatccggca
      301 gatattccgg actacctgaa gttgtccttc ccggagggtt ttaagtggga gcgtgttatg
      361 aatttcgagg acggcggtgt cgtgaccgtt acccaagaca gcagcctgca agatggcgag
      421 ttcatctata aagttaagct gcgtggtacg aacttcccgt ctgatggccc ggtgatgcag
      481 aaaaagacca tgggttggga agcgagcagc gagcgcatgt acccggaaga cggtgcgctg
      541 aagggtgaga tcaaacagcg cctgaagttg aaggacggtg gccactacga tgcagaagta
      601 aaaaccacct ataaggctaa aaaaccggtt cagttgccgg gtgcctataa cgtgaatatt
      661 aagttagaca tcaccagcca taacgaagac tacaccattg tggaacaata tgagcgcgcg
      721 gagggtcgtc acagcaccgg tggtatggat gaactgtaca agtaatgata atagggtacc
      781 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      841 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      901 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      961 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     1021 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     1081 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     1141 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     1201 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1261 tggcactttt cggggaaatg tgtgatcggc acgtaagagg ttccaacttt caccataatg
     1321 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1381 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1441 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1501 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1561 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     1621 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1681 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1741 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1801 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1861 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1921 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     1981 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2041 cgtaacctcg ctaacggatt caccactccg cagacccgcc ataaaacgcc ctgagaagcc
     2101 cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata aaaggcgcct
     2161 gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact gaatgtcacg
     2221 aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc agcgatttgc
     2281 ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt tgtagaggag
     2341 caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt agtgagttat
     2401 acacagggct gggatctatt ctttttatct ttttttattc tttctttatt ctataaatta
     2461 taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt acagagggtc
     2521 tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata cccacaactc
     2581 aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat agtgattaaa
     2641 atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca aatgaactag
     2701 cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc tgtgtggcac
     2761 tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg ttcacttata
     2821 accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta ttagctaaag
     2881 caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt aaaggctttg
     2941 agattttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa ttagttttta
     3001 gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat aatctggaac
     3061 atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta ttaaaagaac
     3121 taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa tttaagttca
     3181 tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg gttttgaaac
     3241 caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat aagcgaggcc
     3301 gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat ctcgtaaccg
     3361 aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc attacatcag
     3421 attcctacct acataacgga ctaagaaaaa cactacacga tgctttaact gcaaaaattc
     3481 agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat gatctcaatg
     3541 gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata ctggctaaat
     3601 acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc aagactaaca
     3661 aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc catatgcaca
     3721 gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt tggtgcattc
     3781 aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt aacacctaat
     3841 agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca cctgagacaa
     3901 cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct gcttatcgaa
     3961 tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa atcatggcaa
     4021 ttctggaaga aatagcgctt tcagccggca aaccggctga agccggatct gcgattctga
     4081 taacaaacta gcaacaccag aacagcccgt ttgcgggcag caaaacccgt acttttggac
     4141 gttccggcgg ttttttgtgg cgagtggtgt tcgggcggtg cgcgcaagat ccattatgtt
     4201 aaacgggcga gtttacatct caaaaccgcc cgcttaacac catcagaaat cctcagcgcg
     4261 attttaagca ccaacccccc cccccgtaac acccaaatcc atactgaaag tggctttgtt
     4321 gaataaatca gatttcgggt aagtctccaa cgccagcaac gcggcctttt tacggttcct
     4381 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4441 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4501 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4561 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4621 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4681 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     4741 cagctatga
//
