LOCUS       Exported                6584 bp DNA     circular SYN 01-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDNA3.1(+)-NEMOs
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2379..2603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2649..3077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3091..3420
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3271..3406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3487..4086
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     polyA_signal    4260..4381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4430..4446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4454..4470
                     /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(4478..4508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4523..4544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4832..5417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5588..6448)
                     /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(6449..6553)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgctgcaga acgagcttgc
      961 tcttaagttg gctggacttg atattaacaa gactggaggg atcactctcg gcatggacga
     1021 gctttacaag atgcatcatc atcatcatca tgtgagcaag ggcgaggagg agaacatggc
     1081 cagcctgcct gctacacatg aactgcacat ttttggaagc atcaacggca tcgacttcga
     1141 catggtgggc cagggcactg gaaatcctaa tgacggatac gaggagctga atctgaagag
     1201 caccatgggc gacctgcagt tcagcccctg gatactggtg cctcatattg gctatggctt
     1261 ccaccaatac ctgccctacc ccgacggaat gagccccttt caagctgcca tggtggatgg
     1321 cagcggatac caagtgcata gaaccatgca gttcgaggac ggcgccagcc ttacagtgaa
     1381 ttatagatat acctacgagg gcagccacat caagggcgag gcacaagtgg aaggcacagg
     1441 atttcccgct gatggccccg tgatgaccaa tagcctgacc gctgaagccc acgaccagct
     1501 gacagaagaa cagatcgccg aattcaagga ggcctttagc ctgttcgaca aggacggcga
     1561 cggcaccatt accacaaaag agctgggcac agtgatgaga agcctgggcc aaaatcccac
     1621 agaagctgag ctgagagtga tgatcatcga ggtggacgcc gacggcgatg gaacactgga
     1681 ttttcctgaa tttctggcca tgatggccag aaagatgaag tacagagaca ccgaggagga
     1741 gatcagagag gccttcggcg tgtttgacaa ggacggcaat ggatacattg gcgccgctga
     1801 actgagacac gtgatgacca atctgggcga gaagctgacc gacgaggagg tgggagaact
     1861 gatcagagag gctgacattg acggcgacgg ccaggtgaat tatgaggagt ttgtgcagat
     1921 gatgaccgcc aagggcggca gcggaggagg aagcagcagc agaagaaaat ggaataaggc
     1981 cggccacgcc gtgagagcca ttggcagact ttctagcatc tacttcgccg actggtgcgt
     2041 gagcaagaag acctgcccta acgacaagac catcgtgagc accttcaagt gggccttcat
     2101 caccgacaac ggcaagagat acagaagcac cgccagaacc acctacacct tcgccaaacc
     2161 catggccgct aactacctga agaaccagcc catgtacgtg ttcagaaaga ccgagctgaa
     2221 gcacagcaag accgagctga acttcaagga gtggcagaag gccttcaccg acgtgatggg
     2281 catggacgag ctgtacaagt aagaattctg cagatatcca gcacagtggc ggccgctcga
     2341 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2401 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2461 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2521 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2581 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2641 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2701 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2761 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2821 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2881 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2941 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3001 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3061 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3241 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3301 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3361 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3421 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3481 tttcgcatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3541 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     3601 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3661 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3721 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     3781 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     3841 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     3901 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     3961 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4021 accgccgacg tcgagtgccc gaaggaccgc gcgacctggt gcatgacccg caagcccggt
     4081 gcctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4141 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4201 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4381 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5341 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5401 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5461 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5521 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5581 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5641 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5701 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5761 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5821 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5881 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5941 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6001 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6061 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6121 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6181 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6241 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6301 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6361 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6421 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6481 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6541 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
