LOCUS       sequence-114545-        5866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry under the control of Mutant T7 Promoter C4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5866)
  AUTHORS   Jones JA, Vernacchio VR, Lachance DM, Lebovich M, Fu L, Shirke AN, 
            Schultz VL, Cress B, Linhardt RJ, Koffas MA
  TITLE     ePathOptimize: A Combinatorial Approach for Transcriptional 
            Balancing of Metabolic Pathways.
  JOURNAL   Sci Rep. 2015 Jun 11;5:11301. doi: 10.1038/srep11301.
  PUBMED    26062452
REFERENCE   2  (bases 1 to 5866)
  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: "Sci Rep."; 
            date: "2015-06-11"; volume: "5:11301. doi"; pages: " 
            10.1038/srep11301"
COMMENT     Sequence Label: pETM6-C4-mCherry
FEATURES             Location/Qualifiers
     source          1..5866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             818..862
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(901..919)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      915..962
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1006..1461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1303..1324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1576..2367,2368..2436))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2199..2218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2437..2529)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2610..3198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3099..3118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3352..3369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3384..3526
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3452..3471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3612..3634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3628..3819)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4391..5473)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5435..5454
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5474..5551)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5757..5776
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactatcaag gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggtttcaaaa ggcgaagaag acaacatggc
      121 gattatcaag gaatttatgc gtttcaaggt ccacatggaa ggcagcgtca atggtcacga
      181 atttgaaatt gaaggcgaag gtgaaggccg tccgtatgaa ggcacccaga cggcaaaact
      241 gaaggtcacc aaaggcggtc cgctgccgtt tgcttgggat attctgtcac cgcaattcat
      301 gtatggttcg aaagcgtacg ttaagcatcc ggccgatatc ccggactatc tgaaactgtc
      361 ctttccggaa ggcttcaaat gggaacgtgt tatgaacttc gaagatggcg gtgtggttac
      421 cgtcacgcag gatagctctc tgcaagacgg tgaatttatt tataaagtga agctgcgcgg
      481 caccaatttc ccgagcgatg gtccggttat gcagaaaaag acgatgggct gggaagcgag
      541 ttccgaacgt atgtacccgg aagacggtgc cctgaaaggc gaaatcaagc agcgcctgaa
      601 actgaaggat ggcggtcact atgacgcaga agtgaaaacc acgtacaagg ctaaaaagcc
      661 ggtccaactg ccgggtgcat acaacgtgaa catcaagctg gatatcacca gccataacga
      721 agactatacg atcgttgaac agtacgaacg tgcagaaggc cgccactcta ccggcggtat
      781 ggatgaactg tacaaataag gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt
      841 tgaacgccag cacatggact cgtctactag tcgcagctta attaacctaa actgctgcca
      901 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      961 tgctagcgaa aggaggagtc gactatatcc ggattggcga atgggacgcg ccctgtagcg
     1021 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1081 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1141 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1201 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1261 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1321 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1381 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1441 aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa aaaggatctt
     1501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatcatgatt
     2461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2521 ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3001 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3241 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3301 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3361 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3421 atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3481 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3541 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3601 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3661 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3721 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3781 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     3841 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3901 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3961 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4021 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4081 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     4141 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     4201 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     4261 caggagcacg atcatgctag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4321 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     4381 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4441 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     4501 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4561 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     4621 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     4681 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     4741 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     4801 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     4861 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     4921 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     4981 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     5041 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     5101 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     5161 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     5221 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     5281 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     5341 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     5401 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     5461 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5521 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     5581 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     5641 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5701 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5761 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5821 cgtccggcgt agcctaggat cgagatcgat ctcgatcccg cgaaat
//
