LOCUS       Exported                6061 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 6061)
  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..6061
                     /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..2211
                     /codon_start=1
                     /label=Tigger1
                     /translation="MASKCSSERKSRTSLTLNQKLEMIKLSEEGMSKAEIGRKLGLLRQ
                     TVSQVVNAKEKFLKEIKSATPVNTRMIRKRNSLIADMEKVLVVWIEDQTSHNIPLSQSL
                     IQSKALTLFNSMKAERGEEAAEEKFEASRGWFMRFKERSHLHNIKVQGEAASADVEAAA
                     SYPEDLAKIIDEGGYTKQQIFNVDETAFYWKKMPSRTFIAREEKSMPGFKASKDRLTLL
                     LGANAAGDFKLKPMLIYHSENPRALKNYAKSTLPVLYKWNNKAWMTAHLFTAWFTEYFK
                     PTVETYCSEKKIPFKILLLIDNAPGHPRALMEMYKEINVVFMPANTTSILQPMDQGVIS
                     TFKSYYLRNTFRKAIAAIDSDSSDGSGQSKLKTFWKGFTILDAIKNIRDSWEEVKISTL
                     TGVWKKLIPTLMDDFEGFKTSVEEVTADVVEIARELELEVEPEDVTELLQSHDKT"
     promoter        complement(2249..2267)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2293..2517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2563..2991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3005..3334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3185..3320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3500..3621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3670..3686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3694..3710
                     /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(3718..3748)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3763..3784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4072..4660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4831..5691)
                     /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(5692..5796)
                     /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 cgagcaaatg tagcagcgag cgcaaaagcc gtactagcct gaccctgaac
      901 cagaaactgg agatgattaa actgagcgaa gagggcatga gcaaggcaga aattggccgc
      961 aaattaggct tactgcgcca aaccgtgagc caagtggtga acgcgaaaga aaagttcctg
     1021 aaggaaatca aaagcgccac ccctgtgaat actcgcatga tccgcaaacg caacagcctg
     1081 attgccgaca tggagaaagt gctggtggtg tggatcgaag atcagaccag ccacaacata
     1141 ccgctgagcc agagcctgat tcagagcaag gccctgaccc tgtttaacag catgaaagcg
     1201 gaacgcggtg aagaagccgc cgaagaaaag tttgaagcga gccgcggctg gtttatgcgc
     1261 tttaaagagc gcagccacct gcataacatt aaagtgcagg gcgaagcggc aagcgcagat
     1321 gttgaagcag cggcgagcta ccctgaagac ctggcgaaga ttattgatga aggcggctat
     1381 accaaacagc agattttcaa cgtggacgag actgcgtttt attggaagaa gatgccgagc
     1441 cgcaccttta ttgcgcgcga ggaaaagagc atgcctggct ttaaggcgag caaagaccgt
     1501 ctgaccttat tactgggcgc gaacgcggca ggtgacttca aactgaagcc gatgctgatc
     1561 tatcacagcg aaaacccgcg cgcgctgaag aattacgcga aaagcaccct gcctgtgctg
     1621 tataaatgga acaacaaggc gtggatgacc gcgcatttat tcaccgcgtg gttcaccgaa
     1681 tattttaagc cgaccgtgga aacctactgc agcgaaaaaa agatcccgtt taagatcctg
     1741 ctgctgattg ataacgcgcc tggtcatcct cgtgcgctga tggaaatgta taaggagatc
     1801 aacgtggtgt tcatgccggc caacaccacc tcaattctgc agcctatgga tcagggcgtg
     1861 atcagcacct tcaaaagcta ctacctgcgc aacaccttcc gcaaagcgat tgccgcgatt
     1921 gatagcgata gcagcgacgg tagcggtcag agcaaactga agaccttttg gaaaggcttc
     1981 accattctgg atgcgattaa aaacattcgc gacagctggg aggaagtgaa aattagcacc
     2041 ctgaccggcg tgtggaaaaa actgattcct accctgatgg acgactttga gggcttcaag
     2101 accagcgtgg aggaagttac cgccgacgtg gttgaaattg cgcgcgagct ggaattagaa
     2161 gtggaaccgg aggatgtgac tgaactgctg cagagccatg ataagacctg atgagcggcc
     2221 gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     2281 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2341 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2401 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2461 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2521 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2581 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2641 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2701 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2761 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2821 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2881 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2941 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3001 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3241 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3301 aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgaaatga ccgaccaagc
     3361 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3421 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3481 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3601 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     3661 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3721 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3781 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3841 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3901 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3961 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5761 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5821 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5881 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5941 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6001 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     6061 t
//
