LOCUS       sequence-9060-ex        4042 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4042)
  TITLE     pTREtight with high copy E. coli origin
REFERENCE   2  (bases 1 to 4042)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pTREtight2-HA_GDP1
FEATURES             Location/Qualifiers
     source          1..4042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    154..235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     179..198
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1485..1511)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1546..1860)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(1549..1573)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1620..1638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1656..1674
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1691..1709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1727..1745
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1763..1781
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1798..1816
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1834..1852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(join(1869..1875,1876..1893,1894..1899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1914..1935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2052..2069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2223..2811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2303..2322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2982..3773,3774..3842))
                     /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     3605..3624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3843..3947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4015..4033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactctc gaggcagtga aaaaaatgct ttatttgtga
      181 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      241 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      301 caagtaaaac ctctacaaat gtggtatggc tgattatgat cctctagaga tatcgtcgac
      361 aagcttatcg atgcggccgc ttaaacacca gcttcgaagt ccttttgagc catgaaaatg
      421 gataaatcaa ccactcttga agagtaacca tattcattat cataccaaga aaggaccttg
      481 aaaaaatggt cgttcaattc aataccggcc ttggcatcaa caatagatga acgtgaatcg
      541 gatgtgaagt cagaggacac aacggcgtct ttggtaacac ccaaaacacc cttcatatcg
      601 ctgcgagatc tttgttctag ggccttcata atgtcatcgt aagaagtttt ctttgctgta
      661 cggaatgtca agtcaaccag ggaaatatta attgttggga ctcttataga cataccggtg
      721 atcttaccat taagttcagg caagattttc cctacagcct tagctgcacc agtagatgaa
      781 ggaatgatat ttccctggca agatctaccg cctctccagt ccttaccacc agaactggta
      841 ccatcgacag tcttttgaga agcagtagtt gcatgaatag ttgtcatcaa ggcttcttcg
      901 ataccgaact catcgtccaa agccttaacc aacggagcca aacagttggt agtacaggag
      961 gcattagaga ccacgtgatc cgtcaatggg ttgtatttaa cgtggttaac accatagacg
     1021 tacattggcg cggtctttga tggagcagta atgataactt ttttgacacc tttatgtcta
     1081 gaggctgtat cgacttcctt gaagacaccg gttgagtcaa ttacataatc gacgttgtag
     1141 gaagcccatg ggatacgctc tggttcccta aaatgagata gagggatatg agccgaaaca
     1201 tggtcatttt gaatgatgat acgttcatcg tcgaattcaa cttcaccacg atacttgccg
     1261 tgagtagaat cgtatttgaa caaataagca gcgtattctg gtgttgtgga tggattattg
     1321 attaatctga ccttaacttc tgggtgcgtc aaagcagcac gtagaaccaa tctaccgatt
     1381 ctaccaaaac cattgatacc aatgttaatt tgagctggct tagaagaaga ttcgtttgtc
     1441 atatcgggca ttgtcgaccg aattcgggcc tccatggcca taagagcgta atctggaaca
     1501 tcgtatgggt acatggatcc ccgggtaccg agctcgaatt ctccaggcga tctgacggtt
     1561 cactaaacga gctctgctta tataggcctc ccaccgtaca cgcctacctc gacatacgtt
     1621 ctctatcact gatagggagt aaactcgaca tacgttctct atcactgata gggataaact
     1681 cgacatacgt tctctatcac tgatagggag taaactcgac atacgttctc tatcactgat
     1741 agggagtaaa ctcgacatac gttctctatc actgataggg agtaaactcg acatcgttct
     1801 ctatcactga tagggagtaa actcgacata cgttctctat cactgatagg gagtaaactc
     1861 gagccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1921 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1981 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2041 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2101 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2161 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2221 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2281 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2341 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2401 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2461 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2521 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2581 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2641 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2701 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2761 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2821 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2881 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2941 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3001 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3061 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3121 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3181 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3241 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3301 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3361 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3421 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3481 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3541 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3601 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3661 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3721 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3781 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3841 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3901 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3961 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4021 cgtatcacga ggccctttcg tc
//
