LOCUS       Exported                5197 bp ds-DNA     circular SYN 24-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5197)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5197
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        228..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        608..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    888
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    900..956
                     /label=GPA_signal sequence
     CDS             957..986
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             996..1274
                     /codon_start=1
                     /label=FRB
                     /translation="MLEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKE
                     TSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISKTR"
     misc_feature    1281..1673
                     /label=hGPA
     misc_feature    1683..1742
                     /label=20GS
     misc_feature    1743..1778
                     /label=114
     misc_feature    1781..1803
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1829..2053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2099..2527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2597..2732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2866..2987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3060..3076
                     /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(3084..3114)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3129..3150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3438..4023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4194..5054)
                     /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(5055..5159)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
       61 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      121 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      181 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      841 ctggcttatc gaaattaata cgactcacta tagggagtcc caagctggct agcgccacca
      901 tgtatggaaa aataatcttt gtattactat tgtcagaaat tgtgagcata tcagcagagc
      961 agaaactcat ctcagaagag gatctgggcg cgcctatgct cgagatgtgg catgaaggcc
     1021 tggaagaggc atctcgtttg tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc
     1081 tggagccctt gcatgctatg atggaacggg gcccccagac tctgaaggaa acatccttta
     1141 atcaggccta tggtcgagat ttaatggagg cccaagagtg gtgcaggaag tacatgaaat
     1201 cagggaatgt caaggacctc ctccaagcct gggacctcta ttatcatgtg ttccgacgaa
     1261 tctcaaagac tagaaccggt ttaagtacca ctgaggtggc aatgcacact tcaacctctt
     1321 cttcagtcac aaagagttac atctcatcac agacaaatga tacgcacaaa cgggacacat
     1381 atgcagccac tcctagagct catgaagttt cagaaatttc tgttagaact gtttaccctc
     1441 cagaagagga aaccggagaa agggtacaac ttgcccatca tttctctgaa ccagagataa
     1501 cactcattat ttttggggtg atggctggtg ttattggaac gatcctctta atttcttacg
     1561 gtattcgccg actgataaag aaaagcccat ctgatgtaaa acctctcccc tcacctgaca
     1621 cagacgtgcc tttaagttct gttgaaatag aaaatccaga gacaagtgat caattaatta
     1681 agggggggtc tggaggcagt ggtgggtctg gtggtggttc tggggggagt gggggttcag
     1741 gcgtgaccgg ctaccggctg ttcgaggaga ttctgtaagc ggccgctcga gtctagaggg
     1801 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1861 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1921 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1981 gtggggcagg acagcaaggg ggaggattgg gaagtcaata gcaggcatgc tggggatgcg
     2041 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2221 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2461 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2521 gcgaattaat tctgtggaat gtgtgtcagt taggctcaat tagtcagcaa ccatagtccc
     2581 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2641 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2701 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     2761 ttgtatatcc attttcggag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     2821 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     2881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     3001 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3961 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4201 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4261 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4321 gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4381 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4441 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4501 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4561 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4621 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4681 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4741 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4801 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4861 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4921 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4981 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5041 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5101 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5161 acatttcccc gaaaagtgcc acctgacgtc gacggat
//
