LOCUS       Exported                3920 bp ds-DNA     circular SYN 12-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3920)
  AUTHORS   hwinn
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 20, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3920
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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     69..88
                     /label=pQE Type III/IV Forward
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             115..117
                     /codon_start=1
                     /label=ATG
                     /translation="M"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             172..423
                     /codon_start=1
                     /label=RTXWT
                     /translation="GSARDDVLIGDAGANVLNGLAGNDVLSGGAGDDVLLGDEGSDLLS
                     GDAGNDDLFGGQGDDTYLFGVGYGHDTIYESGGGHDTIR"
     CDS             172..423
                     /codon_start=1
                     /label=RTXWTM-X2D
                     /translation="GSARDDVLIGDAGDNVLNGLAGDDVLSGGAGDDVLLGDEGDDLLS
                     GDAGDDDLFGGQGDDTYLFGVGYGDDTIYESGGGDDTIR"
     CDS             424..627
                     /codon_start=1
                     /label=C-Cap
                     /translation="INAGADQLWFARQGNDLEIRILGTDDALTVHDWYRDADHRVEIIH
                     AANQAVDQAGIEKLVEAMAQYPD"
     CDS             424..627
                     /codon_start=1
                     /label=C-Cap
                     /translation="INAGADQLWFARQGNDLEIRILGTDDALTVHDWYRDADHRVEIIH
                     AANQAVDQAGIEKLVEAMAQYPD"
     terminator      667..761
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(684..702)
                     /label=pQE Reverse
     CDS             805..1464
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            805..1464
                     /label=CAT/CamR
     terminator      1529..1615
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1532..1575
                     /label=rrnB_T1_terminator
     misc_feature    1770..1910
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2096..2684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2855..3715)
                     /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"
     gene            complement(2855..3715)
                     /label=Ampicillin
     promoter        complement(3716..3820)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(3757..3785)
                     /label=AmpR_promoter
     primer_bind     3829..3846
                     /label=pQE Forward
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc catatggagc tcggcgctag cggttctgcc
      181 agagacgatg tactcattgg tgatgcgggt gataatgttc tcaacggttt agcaggcgat
      241 gacgttctga gtgggggagc tggtgacgac gtattactgg gcgatgaagg agatgatctg
      301 cttagcggag atgctggcga tgatgacttg ttcgggggtc agggcgatga cacgtatttg
      361 tttggagtgg gctatggtga cgacacaatc tatgagtcag ggggtgggga tgacaccatt
      421 cgtatcaatg caggcgcgga tcaactgtgg tttgctcggc agggcaatga tcttgagatc
      481 cgcatactgg gtactgatga tgccctgacc gtccatgatt ggtaccgtga cgcggatcat
      541 cgcgttgaaa taattcacgc cgctaaccaa gcagtggatc aggccggaat tgaaaaatta
      601 gtcgaagcga tggcacagta cccggatgaa ttcactagtc tcgagaagct taattagctg
      661 agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
      721 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag cttggcgaga
      781 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
      841 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
      901 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
      961 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     1021 tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     1081 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     1141 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     1201 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     1261 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     1321 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     1381 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     1441 gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct
     1501 ggggtaatga ctctctagct tgaggcatca aataaaacga aaggctcagt cgaaagactg
     1561 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1621 ctctagagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1681 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1741 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     1801 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     1861 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     1921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2101 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2521 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2701 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2761 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2821 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2881 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2941 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3001 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3061 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3121 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3181 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3241 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3301 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3361 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3421 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3481 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3541 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3601 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3661 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3721 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3781 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3841 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3901 cgaggccctt tcgtcttcac
//
