LOCUS       Exported                5716 bp ds-DNA     circular SYN 22-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5716
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             847..1866
                     /codon_start=1
                     /label=Tc1-1_PS
                     /translation="MKFKELSIDLRDRIVSRHRSGEGYRKISAALKVPMSTVASIIRTW
                     KKFGTTRTLPRVGRPARLSDRGRRALVREVTRNPMVTLTELQRCSMKRGEPSRRTTISA
                     ALHKSGLFGRVARRKPLLSKKHMTARLEFAKKHLKDSQTMRNKILWSDETKIELFGLNA
                     KRHVWRKPGTAHHLANTIPTVKHGGGSIMLWGCFSAGGTGRRVRIEGKMNAAMYRDILD
                     ENMLQSALDLRLGRRFIFQQDNDPKHTAEITKEWLRDDSVNVLEWPSQSPDLNPIEHLW
                     RDLKMAVHRRSPSNLMELERFCKEEWEKLPRNRCATLVESYPRRLEAVIAAKGASTKY"
     promoter        complement(1904..1922)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1948..2172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2218..2646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2660..2989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2840..2975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3155..3276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3325..3341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3349..3365
                     /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)."
     promoter        complement(3373..3403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3418..3439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3727..4315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4486..5346)
                     /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(5347..5451)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacgtc cgcgcagggc aatagcagag ctctctggct
      721 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      781 ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcgcc
      841 gccaccatga agttcaagga gctgagcatt gatctgcgcg atcgcatcgt tagccgtcat
      901 cgtagcggtg aaggctatcg caaaattagc gcggcgctga aggttcctat gagcaccgtg
      961 gcgagcataa tacgcacctg gaagaaattt ggcaccaccc gtaccttacc tcgtgttggc
     1021 cgtcctgcgc gtttaagcga tcgtggtcgt cgtgcgttag ttcgtgaggt gacccgcaat
     1081 cctatggtga ccctgaccga attacagcgc tgcagcatga aacgtggcga acctagccgc
     1141 cgtactacca ttagcgcggc gttacacaag agcggcctgt ttggtcgtgt ggcgcgtcgt
     1201 aaacctctgc tgagcaaaaa acatatgacc gcccgcctgg agttcgcgaa gaaacatctg
     1261 aaggatagcc agaccatgcg caacaaaatc ctgtggagcg atgaaaccaa aatcgagctg
     1321 tttggcctga acgcgaaacg ccatgtgtgg cgcaaacctg gtaccgcaca tcatctggcg
     1381 aacaccattc cgaccgtgaa acatggtggc ggcagcatta tgctgtgggg ctgcttttca
     1441 gcaggtggta ccggtcgtcg tgttcgtatt gaaggcaaaa tgaacgccgc catgtaccgc
     1501 gatattctgg atgagaacat gctgcagagc gcactggact tacgtttagg ccgtcgcttt
     1561 atcttccagc aggataacga ccctaagcat accgccgaga tcaccaaaga gtggctgcgc
     1621 gatgatagcg tgaacgtgct ggaatggcct agccaaagcc ctgatctgaa cccgattgaa
     1681 catctgtggc gcgacctgaa aatggcagtt caccgtcgct cacctagcaa cctgatggaa
     1741 ctggaacgct tctgcaaaga ggaatgggaa aagctgccgc gtaatcgttg tgcgaccctg
     1801 gttgagtcat atcctcgtcg cctggaagca gttatagcag cgaaaggcgc cagcaccaag
     1861 tattgatgag cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     1921 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1981 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2041 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2101 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2161 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2221 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2281 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2341 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2401 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2461 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2521 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2581 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2641 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2701 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2761 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2821 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2881 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2941 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccga
     3001 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3061 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3121 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3181 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3241 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3301 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3361 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3421 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3481 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3541 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4141 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4621 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5341 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5401 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5461 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     5521 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     5581 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     5641 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     5701 gccagatata cgcgtt
//
