LOCUS       sequence-181154-        2932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2932)
  TITLE     pTREtight with high copy E. coli origin
REFERENCE   2  (bases 1 to 2932)
  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"
COMMENT     Sequence Label: pTREtight2
FEATURES             Location/Qualifiers
     source          1..2932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..25)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    72..90
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    108..126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    143..161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    179..197
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    215..233
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    250..268
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    286..304
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(join(321..327,328..345,346..351))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    366..387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(504..521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(675..1263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(755..774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1434..2225,2226..2294))
                     /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     2057..2076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2295..2399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2467..2485
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2523..2545)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    2648..2729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2673..2692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(join(2930..2932,1..312))
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
ORIGIN
        1 gatctgacgg ttcactaaac gagctctgct tatataggcc tcccaccgta cacgcctacc
       61 tcgacatacg ttctctatca ctgataggga gtaaactcga catacgttct ctatcactga
      121 tagggataaa ctcgacatac gttctctatc actgataggg agtaaactcg acatacgttc
      181 tctatcactg atagggagta aactcgacat acgttctcta tcactgatag ggagtaaact
      241 cgacatcgtt ctctatcact gatagggagt aaactcgaca tacgttctct atcactgata
      301 gggagtaaac tcgagccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      361 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      421 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      481 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      541 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      601 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1141 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1201 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1381 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1441 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1501 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1561 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1621 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1681 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1741 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1801 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1861 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1921 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1981 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2041 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2101 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2161 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2221 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2281 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2341 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2401 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2461 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     2521 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2581 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     2641 tctcgaggca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2701 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2761 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2821 tggctgatta tgatcctcta gagatatcgt cgacaagctt atcgatgcgg ccgcgctagc
     2881 acgcgtcagc tgactagagg atccccgggt accgagctcg aattctccag gc
//
