LOCUS       Exported                3911 bp ds-DNA     circular SYN 11-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3911)
  AUTHORS   Stanford University
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 20, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3911
                     /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..414
                     /codon_start=1
                     /label=RTXMCR-TLY-X2S
                     /translation="GGAGSDTLYGGAGSDTLYGGAGSDTLYGGAGSDTLYGGAGSDTLY
                     GGAGSDTLYGGAGSDTLYGGAGSDTLYGGAGSDTLY"
     CDS             415..618
                     /codon_start=1
                     /label=C-Cap
                     /translation="INAGADQLWFARQGNDLEIRILGTDDALTVHDWYRDADHRVEIIH
                     AANQAVDQAGIEKLVEAMAQYPD"
     terminator      658..752
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(675..693)
                     /label=pQE Reverse
     gene            796..1455
                     /label=CAT/CamR
     CDS             796..1455
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1520..1606
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1523..1566
                     /label=rrnB_T1_terminator
     misc_feature    1761..1901
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2087..2675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(2846..3706)
                     /label=Ampicillin
     CDS             complement(2846..3706)
                     /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"
     promoter        complement(3707..3811)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(3748..3776)
                     /label=AmpR_promoter
     primer_bind     3820..3837
                     /label=pQE Forward
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc catatggagc tcggcgctag cggcggtgca
      181 ggaagtgata cactttacgg cggagcgggc agtgacacgt tgtacggtgg ggcgggttcc
      241 gacactttat acggcggagc aggttcagac actctttacg gtggtgcagg atcagacact
      301 ctctatggcg gggccggctc tgacaccttg tacggcggag ctggttcaga cacgttatat
      361 ggcggcgcgg ggtcagacac actttatggc ggggcaggga gcgacacact gtacattaac
      421 gcgggagccg accagctgtg gttcgcgcga caaggtaatg acttagagat acgtatcctg
      481 gggacagatg acgcacttac ggtgcatgac tggtatagag acgcggacca tcgtgtcgag
      541 ataatacacg cggccaacca ggcggtagac caggccggta ttgagaagct ggtcgaggcg
      601 atggcacaat accccgacga attcactagt ctcgagaagc ttaattagct gagcttggac
      661 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
      721 gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga
      781 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
      841 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
      901 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
      961 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
     1021 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1081 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1141 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1201 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1261 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     1321 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     1381 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     1441 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     1501 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1561 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cctctagagc
     1621 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     1681 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1741 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     1801 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     1861 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     1921 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1981 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2041 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2101 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2161 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2221 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2281 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2341 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2401 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2461 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2521 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2581 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2641 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2701 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2761 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2821 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2881 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2941 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3001 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3061 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3121 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3181 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3241 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3301 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3361 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3421 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3481 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3541 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3601 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3661 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3721 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3781 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3841 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3901 ttcgtcttca c
//
