LOCUS       sequence-444695-        5255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with mCherry integrated in the multiple 
            cloning site (TetR). 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 5255)
  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 5255)
  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: pHBJT032
FEATURES             Location/Qualifiers
     source          1..5255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..31
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15..31
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(823..840)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(823..839)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(832..854)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1048..1067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1167..1189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1227..1245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1317..1322,1323..1339,1340..1345)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1393..2583
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1745..1764)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      2932..3154
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3202..4152
                     /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     5038..5055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5172..5193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5208..5213,5214..5231,5232..5238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(5246..5255,1..7)
                     /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     join(5251..5255,1..18)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
       61 ggtcgactct agaggatccc cgggatggta tcaaaaggag aggaagataa tatggcgatc
      121 atcaaagagt tcatgcgttt taaagtgcac atggaaggct ccgtgaacgg tcacgagttc
      181 gaaattgagg gcgagggcga gggccgtccg tatgaaggca cgcaaactgc taaactcaag
      241 gtcacgaaag gcggaccact gccttttgcc tgggacatcc tgtctccgca gtttatgtac
      301 ggcagcaaag cgtacgttaa acatccggca gatattccgg actacctgaa gttgtccttc
      361 ccggagggtt ttaagtggga gcgtgttatg aatttcgagg acggcggtgt cgtgaccgtt
      421 acccaagaca gcagcctgca agatggcgag ttcatctata aagttaagct gcgtggtacg
      481 aacttcccgt ctgatggccc ggtgatgcag aaaaagacca tgggttggga agcgagcagc
      541 gagcgcatgt acccggaaga cggtgcgctg aagggtgaga tcaaacagcg cctgaagttg
      601 aaggacggtg gccactacga tgcagaagta aaaaccacct ataaggctaa aaaaccggtt
      661 cagttgccgg gtgcctataa cgtgaatatt aagttagaca tcaccagcca taacgaagac
      721 tacaccattg tggaacaata tgagcgcgcg gagggtcgtc acagcaccgg tggtatggat
      781 gaactgtaca agtaatgata atagggtacc gagctcgaat tcactggccg tcgttttaca
      841 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
      901 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgca
      961 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1021 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     1081 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     1141 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     1201 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     1261 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgatgtttga
     1321 cagcttatca tcgataagct ttaatgcggt agtttatcac agttaaattg ctaacgcagt
     1381 caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac cgtcaccctg
     1441 gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc
     1501 cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa
     1561 tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg
     1621 ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg
     1681 tggatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     1741 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     1801 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     1861 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     1921 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc
     1981 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2041 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     2101 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     2161 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     2221 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     2281 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     2341 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     2401 caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg
     2461 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     2521 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     2581 tgacctcgct aacggattca ccactccgca gacccgccat aaaacgtcct gagaagcccg
     2641 tgacgggctt ttcttgtatt atgggtagtt tccttgcatg aatccataaa aggcgcctgt
     2701 agtgccattt acccccattc actgccagag ccgtgagcgc agcgaactga atgtcacgaa
     2761 aaagacagcg actcaggtgc ctgatggtcg gagacaaaag gaatattcag cgatttgccc
     2821 gagcttgcga gggtgctact taagccttta gggttttaag gtctgttttg tagaggagca
     2881 aacagcgttt gcgacatcct tttgtaatac tgcggaactg actaaagtag tgagttatac
     2941 acagggctgg gatctattct ttttatcttt ttttattctt tctttattct ataaattata
     3001 accacttgaa tataaacaaa aaaaacacac aaaggtctag cggaatttac agagggtcta
     3061 gcagaattta caagttttcc agcaaaggtc tagcagaatt tacagatacc cacaactcaa
     3121 aggaaaagga ctagtaatta tcattgacta gcccatctca attggtatag tgattaaaat
     3181 cacctagacc aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg
     3241 attagtcgct atgacttaac ggagcatgaa accaagctaa ttttatgctg tgtggcacta
     3301 ctcaacccca cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt cacttataac
     3361 caatacgctc agatgatgaa catcagtagg gaaaatgctt atggtgtatt agctaaagca
     3421 accagagagc tgatgacgag aactgtggaa atcaggaatc ctttggttaa aggctttgag
     3481 attttccagt ggacaaacta tgccaagttc tcaagcgaaa aattagaatt agtttttagt
     3541 gaagagatat tgccttatct tttccagtta aaaaaattca taaaatataa tctggaacat
     3601 gttaagtctt ttgaaaacaa atactctatg aggatttatg agtggttatt aaaagaacta
     3661 acacaaaaga aaactcacaa ggcaaatata gagattagcc ttgatgaatt taagttcatg
     3721 ttaatgcttg aaaataacta ccatgagttt aaaaggctta accaatgggt tttgaaacca
     3781 ataagtaaag atttaaacac ttacagcaat atgaaattgg tggttgataa gcgaggccgc
     3841 ccgactgata cgttgatttt ccaagttgaa ctagatagac aaatggatct cgtaaccgaa
     3901 cttgagaaca accagataaa aatgaatggt gacaaaatac caacaaccat tacatcagat
     3961 tcctacctac ataacggact aagaaaaaca ctacacgatg ctttaactgc aaaaattcag
     4021 ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa gtaagtatga tctcaatggt
     4081 tcgttctcat ggctcacgca aaaacaacga accacactag agaacatact ggctaaatac
     4141 ggaaggatct gaggttctta tggctcttgt atctatcagt gaagcatcaa gactaacaaa
     4201 caaaagtaga acaactgttc accgttacat atcaaaggga aaactgtcca tatgcacaga
     4261 tgaaaacggt gtaaaaaaga tagatacatc agagctttta cgagtttttg gtgcattcaa
     4321 agctgttcac catgaacaga tcgacaatgt aacagatgaa cagcatgtaa cacctaatag
     4381 aacaggtgaa accagtaaaa caaagcaact agaacatgaa attgaacacc tgagacaact
     4441 tgttacagct caacagtcac acatagacag cctgaaacag gcgatgctgc ttatcgaatc
     4501 aaagctgccg acaacacggg agccagtgac gcctcccgtg gggaaaaaat catggcaatt
     4561 ctggaagaaa tagcgctttc agccggcaaa ccggctgaag ccggatctgc gattctgata
     4621 acaaactagc aacaccagaa cagcccgttt gcgggcagca aaacccgtac ttttggacgt
     4681 tccggcggtt ttttgtggcg agtggtgttc gggcggtgcg cgcaagatcc attatgttaa
     4741 acgggcgagt ttacatctca aaaccgcccg cttaacacca tcagaaatcc tcagcgcgat
     4801 tttaagcacc aacccccccc cgtaacaccc aaatccatac tgaaagtggc tttgttgaat
     4861 aaatcagatt tcgggtaagt ctccaacgcc agcaacgcgg cctttttacg gttcctggcc
     4921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5041 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5101 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5161 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     5221 cttccggctc gtatgttgtg tggaattgtg agcgg
//
