LOCUS       Exported                6556 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 6556)
  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..6556
                     /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..2706
                     /codon_start=1
                     /label=Tigger2f
                     /translation="MGPIRSATSDAGSAPKKQRKVMTLQEKVELLDMYHRLRSAAAVAR
                     HFKINESSVRTTVKKEKEIREAVAAATPAGAKTLHFLRNTFLSRIENAAFMWVQDCYKK
                     GIPIDSNMIREKAKSLYDNLKQKEGEGSKAGEFNASKGWFDNFRKRFGLKNVKITGEAA
                     SADQEAADEFPDAIKKIIEEKGYLPEQVFNADESALFWRKKMPQRTFISKEEKRAPGFK
                     AGRDRLTLLFCANAVGFMIRTALIYKAANPRALKGKDKHQLPVFWLYNKKAWTTRTLFL
                     DWFHRCFVPEVRKYLASKGLPFKVLLILDNAPGHPEPHEFNTEGVEVVYLPPNTTSLIQ
                     PLDQGVIRTFKAHYTRYSMERIVNAMEENPDRENIMKVWKDYTIEDAIIVIEKAMKAIK
                     PETINSCWRKLCPDVVHDFTGFTTEPIKEIMKEIVDMAKKVGGEGFQDMDLGEIQELID
                     TTPEELTEDDLMEMSASEPVPDDEEEDVEEAVPENKLTLDNLAEGFRLFKTAFDFFYDM
                     DPSMIRALKLKQTVEEGLVPYRNIFREMKKQKSQTEITMYFHKVTLSVPASPASPSTSS
                     TSSASATPETARPTPPLPPPPQPTQREDDEDEDLYDDPLPLNE"
     promoter        complement(2744..2762)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2788..3012
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3058..3486
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3500..3829
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3680..3815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3995..4116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4165..4181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4189..4205
                     /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(4213..4243)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4258..4279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4567..5155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5326..6186)
                     /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(6187..6291)
                     /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 gccaccatgg gtcctattcg tagcgcaacc agcgatgcag gtagcgcgcc taaaaaacag
      901 cgcaaagtga tgaccctgca ggaaaaggtg gagctgctgg acatgtatca tcgcctgcgt
      961 agcgcagcag cagttgcacg ccactttaag attaacgaaa gcagcgtgcg caccaccgtg
     1021 aaaaaagaga aagaaatccg cgaagcggtt gcagcagcaa cccctgcagg tgcgaaaacc
     1081 ttacatttcc tgcgcaacac ctttctgagc cgcattgaaa acgccgcctt tatgtgggtg
     1141 caggactgct ataaaaaagg cattccgatt gacagcaaca tgattcgcga gaaggcgaaa
     1201 agcctgtatg acaacctgaa acagaaagag ggcgaaggta gcaaagcggg cgagttcaac
     1261 gcaagcaaag gctggttcga taactttcgc aaacgctttg gcctgaaaaa cgtgaaaatc
     1321 accggcgaag cagcgtcagc agatcaagaa gcggccgacg aatttcctga tgcgatcaag
     1381 aagattattg aggagaaggg ctacttaccg gaacaggtgt ttaacgcgga cgaaagcgcg
     1441 ctgttttggc gcaaaaagat gccgcagcgc accttcatta gcaaagaaga aaagcgcgcg
     1501 cctggtttca aagcaggccg cgatcgctta accctgctgt tttgcgcaaa cgccgtgggc
     1561 tttatgattc gcaccgcgct gatttataaa gcggccaacc ctcgtgcact gaaaggcaaa
     1621 gataagcatc agctgcctgt gttttggctg tataacaaga aggcctggac cacccgtact
     1681 ctgtttctgg actggttcca tcgctgcttt gtgcctgagg tgcgcaaata tctggcgagc
     1741 aaaggcctgc cgtttaaagt gctgctgatt ctggataacg cgcctggtca tccggagcct
     1801 catgaattta acaccgaggg cgtggaggtt gtgtacttac cgcctaacac caccagcctg
     1861 atccagcctc tggatcaggg tgtgatccgc acctttaagg cccattacac ccgctacagc
     1921 atggaacgca tcgtgaacgc gatggaagaa aacccggatc gcgagaacat tatgaaagtg
     1981 tggaaggact ataccattga ggacgcgatc attgtgatcg aaaaggcgat gaaggcgatc
     2041 aaacctgaaa ccatcaacag ctgctggcgc aaactgtgcc ctgatgtggt gcatgatttc
     2101 accggcttta ccaccgaacc gatcaaagaa atcatgaagg agatcgtgga catggccaaa
     2161 aaagtgggcg gcgaaggctt ccaggatatg gatctgggcg aaattcagga actgattgat
     2221 accaccccgg aagaactgac cgaagatgac ctgatggaga tgagcgccag cgaacctgtt
     2281 ccggacgatg aagaagaaga cgtggaagaa gccgtgccgg agaataagct gaccctggat
     2341 aacctggccg aaggctttcg cctgtttaag accgcgttcg acttttttta tgacatggac
     2401 cctagcatga tccgcgcgct gaaactgaaa cagaccgtgg aggagggctt agttccgtat
     2461 cgcaacattt tccgcgagat gaaaaagcag aaaagccaga ccgagatcac catgtacttc
     2521 cataaagtga ccctgagcgt tcctgcgtca cctgcgagcc cttcaactag cagcacctca
     2581 agcgcatcag cgactcctga aaccgcacgt cctacccctc ctttacctcc tcctcctcaa
     2641 cctacccagc gcgaagatga tgaagatgaa gacctgtacg atgatccgct gcctctgaac
     2701 gagtaatgag cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     2761 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2821 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2881 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2941 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3001 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3061 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3121 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3181 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3241 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3301 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3361 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3421 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3481 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3661 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3721 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3781 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccga
     3841 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3901 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3961 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4021 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4081 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4141 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4201 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4261 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4321 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4381 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4441 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4981 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5281 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5341 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5401 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5461 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5521 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5581 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5641 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5701 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5761 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5821 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5881 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5941 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6001 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6061 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6121 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6181 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6301 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     6361 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     6421 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     6481 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     6541 gccagatata cgcgtt
//
