LOCUS       sequence-271203-        3784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible circular permutation of GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3784)
  AUTHORS   Menacho-Melgar R, Ye Z, Moreb EA, Yang T, Efromson JP, Decker JS, 
            Wang R, Lynch MD
  TITLE     Scalable, two-stage, autoinduction of recombinant protein expression
            in E. coli utilizing phosphate depletion.
  JOURNAL   Biotechnol Bioeng. 2020 May 22. doi: 10.1002/bit.27440.
  PUBMED    32441815
REFERENCE   2  (bases 1 to 3784)
  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: "Biotechnol 
            Bioeng. 2020 May 22. doi: 10.1002/bit.27440."
COMMENT     Sequence Label: pTKhc-yibDp-GFP-cp6
FEATURES             Location/Qualifiers
     source          1..3784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      164..235
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      251..278
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             571..593
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             613..630
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             634..657
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     terminator      1494..1540
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1688..1704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1688..1704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1701..1723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2600..3415)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2600..3415)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3323..3342
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3407..3426
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3536..3552
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3536..3552
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3553..3599
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3713..3735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3727..3744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3728..3744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tccgcctgac ctatatcaag cttcgtccat tgcatgaggc tcgtcctgaa tgatatcaag
       61 cttcgtccat tgcatgaggc tcgtcctgaa tgatatcaag cttcgtccat tgcatgaggc
      121 tcgtcctgaa tgatatcaag cttgaattcg tttgcccagg catcaaataa aacgaaaggc
      181 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
      241 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatacacagc taacaccacg
      301 tcgtccctat ctgctgccct aggtctatga gtggttgctg gataacgtgc gtaattgtgc
      361 tgatctctta tatagctgct ctcattatct ctctaccctg aagtgactct ctcacctgta
      421 aaaataatat ctcacaggct taatagtttc ttaatacaaa gcctgtaaaa cgtcaggata
      481 acttctatat tcagggagac cacaacggtt tccctctaca aataattttg tttaactttg
      541 agaccacaac ggtttcccac tagaaataat tttgtttaac tttaagaagg agatatacat
      601 atgggcagca gccatcacca tcatcaccac agcctggaag ttctgttcca ggggcccgga
      661 tccatgaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
      721 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
      781 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
      841 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
      901 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaaggtgg taccggtggt
      961 tctatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     1021 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     1081 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact
     1141 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     1201 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     1261 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     1321 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     1381 aagctggaat acaatctcga gtaactcgag tctggtaaaa ctagcattcg acctagcata
     1441 accccgcggg gcctcttcgg gggtctcgcg gggttttttg ctgaaagaag cttcaaataa
     1501 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcgctgcggc
     1561 cgggtcaggt atgatttaaa tggtcagtaa cgggtcttga ggggtttttt gcaatgggtt
     1621 catcccgtgg gaggctaggt ggaggctcag tgatgataag tctgcgatgg tggatgcatg
     1681 tgtcatggtc atagctgttt cctgtgtgaa attgttatcc gctcagaggg cacaatccta
     1741 ttccgcgcta tccgacaatc tccaagacat taggtggagt tcagttcggc gtatggcata
     1801 tgtcgctgga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2461 tcaagaagat cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt
     2521 cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg tccgctctgc cagtgttaca
     2581 accaattaac aaattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     2641 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     2701 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     2761 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     2821 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
     2881 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     2941 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     3001 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     3061 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     3121 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     3181 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     3241 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     3301 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     3361 tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg ctcataacac
     3421 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     3481 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttccccc gccgctctag
     3541 aactagtgga tccaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3601 tgttgtttgt cgcattatac gagacgtcca ggttgggata cctgaaacaa aacccatcgt
     3661 acggccaagg aagtctccaa taactgtgat ccaccacaag cgccagggtt ttcccagtca
     3721 cgacgttgta aaacgacggc cagtcatgca taatccgcac gcatctggaa taaggaagtg
     3781 ccat
//
