LOCUS       sequence-323567-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Theta replicating shuttle vector for E. coli and gram positive 
            bacteria. High copy number in gram positives. Erythromycin 
            resistance, recommended to use BHI agar plates for E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  AUTHORS   O'Sullivan DJ, Klaenhammer TR
  TITLE     High- and low-copy-number Lactococcus shuttle cloning vectors with 
            features for clone screening.
  JOURNAL   Gene. 1993 Dec 31;137(2):227-31.
  PUBMED    8299952
REFERENCE   2  (bases 1 to 6721)
  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: "Gene."; 
            date: "1993-12-31"; volume: "137(2)"; pages: "227-31"
COMMENT     Sequence Label: pTRKH2
FEATURES             Location/Qualifiers
     source          1..6721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             212..454
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSISRSRARESLAAGRRIPGNSLAVVLQRRDWENPGVTQ
                     LNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     primer_bind     complement(295..312)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(295..311)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(304..326)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5426..5445
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5512..5614)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6006..6023)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6140..6685)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6220..6239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgaatt catcgatatc
      241 tagatctcga gctcgcgaaa gcttggctgc aggtcgacgg atccccggga attcactggc
      301 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
      361 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      421 ccaacagttg cgcagcctga atggcgaatg ctagaatcga tacgattttg aagtggcaac
      481 agataaaaaa aagcagttta aaattgttgc tgaactttta aaacaagcaa atacaatcat
      541 tgtcgcaaca gatagcgaca gagaaggcga aaacattgcc tggtcgatca ttcataaagc
      601 aaatgccttt tctaaagata aaacgtataa aagactatgg atcaatagtt tagaaaaaga
      661 tgtgatccgt agcggttttc aaaatttgca accaggaatg aattactatc ccttttatca
      721 agaagcgcac aaaaagaaaa acgaaatgat acaccaatca gtgcaaaaaa agatataatg
      781 ggagataaga cggttcgtgt tcgtgctgac ttgcaccata tcataaaaat cgaaacagca
      841 aagaatggcg gaaacgtaaa agaagttatg gaaataagac ttagaagcaa acttaagagt
      901 gtgttgatag tgcagtatct taaaattttg tataatagga attgaagtta aattagatgc
      961 taaaaatttg taattaagaa ggagtgatta catgaacaaa aatataaaat attctcaaaa
     1021 ctttttaacg agtgaaaaag tactcaacca aataataaaa caattgaatt taaaagaaac
     1081 cgataccgtt tacgaaattg gaacaggtaa agggcattta acgacgaaac tggctaaaat
     1141 aagtaaacag gtaacgtcta ttgaattaga cagtcatcta ttcaacttat cgtcagaaaa
     1201 attaaaactg aatactcgtg tcactttaat tcaccaagat attctacagt ttcaattccc
     1261 taacaaacag aggtataaaa ttgttgggag tattccttac catttaagca cacaaattat
     1321 taaaaaagtg gtttttgaaa gccatgcgtc tgacatctat ctgattgttg aagaaggatt
     1381 ctacaagcgt accttggata ttcaccgaac actagggttg ctcttgcaca ctcaagtctc
     1441 gattcagcaa ttgcttaagc tgccagcgga atgctttcat cctaaaccaa aagtaaacag
     1501 tgtcttaata aaacttaccc gccataccac agatgttcca gataaatatt ggaagctata
     1561 tacgtacttt gtttcaaaat gggtcaatcg agaatatcgt caactgttta ctaaaaatca
     1621 gtttcatcaa gcaatgaaac acgccaaagt aaacaattta agtaccgtta cttatgagca
     1681 agtattgtct atttttaata gttatctatt atttaacggg aggaaataat tctatgagtc
     1741 gcttttgtaa atttggaaag ttacacgtta ctaaagggaa tgtagataaa ttattaggta
     1801 tactactgac agcttccaag gagctaaaga ggtccctagc gctcttatca tggggaagct
     1861 cggatcatat gcaagacaaa ataaactcgc aacagcactt ggagaaatgg gacgaatcga
     1921 gaaaaccctc tttacgctgg attacatatc taataaagcc gtaaggagac gggttcaaaa
     1981 aggtttaaat aaaggagaag caatcaatgc attagctaga attatatttt ttggacaacg
     2041 tggagaattt agagaacgtg ctctccaaga ccagttacaa agagctagtg cactaaacat
     2101 aattattaac gctataagtg tgtggaacac tgtatatatg gaaaaagccg tagaagaatt
     2161 aaaagcaaga ggagaattta gagaagattt aatgccatat gcgtggccgt taggatggga
     2221 acatatcaat tttcttggag aatacaaatt tgaaggatta catgacactg ggcaaatgaa
     2281 tttacgtcct ttacgtataa aagagccgtt ttattcttaa tataacggct ctttttatag
     2341 aaaaaatcct tagcgtggtt tttttccgaa atgctggcgg taccccaaga attagaaatg
     2401 agtagatcaa attattcacg aatagaatca ggaaaatcag atccaaccat aaaaacacta
     2461 gaacaaattg caaagttaac taactcaacg ctagtagtgg atttaatccc aaatgagcca
     2521 acagaaccag aaccagaaac agaatcagaa caagtaacat tggatttaga aatggaagaa
     2581 gaaaaaagca atgacttcgt gtgaataatg cacgaaatcg ttgcttattt tttttaaaag
     2641 cggtatacta gatataacga aacaacgaac tgaatagaaa cgaaaaaaga gccatgacac
     2701 atttataaaa tgtttgacga cattttataa atgcatagcc cgataagatt gccaaaccaa
     2761 cgcttatcag ttagtcagat gaactcttcc ctcgtaagaa gttatttaat taactttgtt
     2821 tgaagacggt atataaccgt actatcatta tatagggaaa tcagagagtt ttcaagtatc
     2881 taagctactg aatttaagaa ttgttaagca atcaatcgga aatcgtttga ttgctttttt
     2941 tgtattcatt tatagaaggt ggagtttgta tgaatcatga tgaatgtaaa acttatataa
     3001 aaaatagttt attggagata agaaaattag caaatatcta tacactagaa acgtttaaga
     3061 aagagttaga aaagagaaat atctacttag aaacaaaatc agataagtat ttttcttcgg
     3121 agggggaaga ttatatatat aagttaatag aaaataacaa aataatttat tcgattagtg
     3181 gaaaaaaatt gacttataaa ggaaaaaaat ctttttcaaa acatgcaata ttgaaacagt
     3241 tgaatgaaaa agcaaaccaa gttaattaaa caacctattt tataggattt ataggaaagg
     3301 agaacagctg aatgaatatc ccttttgttg tagaaactgt gcttcatgac ggcttgttaa
     3361 agtacaaatt taaaaatagt aaaattcgct caatcactac caagccaggt aaaagcaaag
     3421 gggctatttt tgcgtatcgc tcaaaatcaa gcatgattgg cggtcgtggt gttgttctga
     3481 cttccgagga agcgattcaa gaaaatcaag atacatttac acattggaca cccaacgttt
     3541 atcgttatgg aacgtatgca gacgaaaacc gttcatacac gaaaggacat tctgaaaaca
     3601 atttaagaca aatcaatacc ttctttattg attttgatat tcacacggca aaagaaacta
     3661 tttcagcaag cgatatttta acaaccgcta ttgatttagg ttttatgcct actatgatta
     3721 tcaaatctga taaaggttat caagcatatt ttgttttaga aacgccagtc tatgtgactt
     3781 caaaatcaga atttaaatct gtcaaagcag ccaaaataat ttcgcaaaat atccgagaat
     3841 attttggaaa gtctttgcca gttgatctaa cgtgtaatca ttttggtatt gctcgcatac
     3901 caagaacgga caatgtagaa ttttttgatc ctaattaccg ttattctttc aaagaatggc
     3961 aagattggtc tttcaaacaa acagataata agggctttac tcgttcaagt ctaacggttt
     4021 taagcggtac agaaggcaaa aaacaagtag atgaaccctg gtttaatctc ttattgcacg
     4081 aaacgaaatt ttcaggagaa aagggtttaa tagggcgtaa taacgtcatg tttaccctct
     4141 ctttagccta ctttagttca ggctattcaa tcgaaacgtg cgaatataat atgtttgagt
     4201 ttaataatcg attagatcaa cccttagaag aaaaagaagt aatcaaaatt gttagaagtg
     4261 cctattcaga aaactatcaa ggggctaata gggaatacat taccattctt tgcaaagctt
     4321 gggtatcaag tgatttaacc agtaaagatt tatttgtccg tcaagggtgg tttaaattca
     4381 agaaaaaaag aagcgaacgt caacgtgttc atttgtcaga atggaaagaa gatttaatgg
     4441 cttatattag cgaaaaaagc gatgtataca agccttattt agtgacgacc aaaaaagaga
     4501 ttagagaagt gctaggcatt cctgaacgga cattagataa attgctgaag gtactgaagg
     4561 cgaatcagga aattttcttt aagattaaac caggaagaaa tggtggcatt caacttgcta
     4621 gtgttaaatc attgttgcta tcgatcatta aagtaaaaaa agaagaaaaa gaaagctata
     4681 taaaggcgct gacaaattct tttgacttag agcatacatt cattcaagag actttaaaca
     4741 agctagcaga acgccctaaa acggacacac aactcgattt gtttagctat gatacaggct
     4801 gaaaataaaa cccgcactat gccattacat ttatatctat gatacgtgtt tgttttttct
     4861 ttgctgttta gcgaatgatt agcagaaata tacagagtaa gattttaatt aattattagg
     4921 gggagaagga gagagtagcc cgaaaacttt tagttggctt ggactgaacg aagtgaggga
     4981 aaggctacta aaacgtcgag gggcagtgag agcgaagcga acacttgatt ttttaatttt
     5041 ctatctttta taggtcatta gagtatactt atttgtccta taaactattt agcagcataa
     5101 tagatttatt gaataggtca tttaagttga gcatattaga ggaggaaaat cttggagaaa
     5161 tatttgaaga acccgattac atggattgga ttagttcttg tggttacgtg gtttttaact
     5221 aaaagtagtg aatttttgat ttttggtgtg tgtgtcttgt tgttagtatt tgctagtcaa
     5281 agtgattaaa tagaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     5341 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     5401 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     5461 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     5521 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     5581 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
     5641 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
     5701 cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tactgattta
     5761 gtgtatgatg gtgtttttga ggtgctccag tggcttctgt ttctatcagc tgtccctcct
     5821 gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc
     5881 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     5941 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     6001 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     6061 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     6121 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     6181 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6241 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     6301 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     6361 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     6421 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     6481 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     6541 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     6601 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     6661 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta
     6721 g
//
