LOCUS       Exported                5434 bp ds-DNA     circular SYN 18-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5434)
  AUTHORS   Brett Stringer
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5434
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     133..152
                     /label=BWS533
     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"
     primer_bind     926..945
                     /label=BWS335
     misc_feature    958..1044
                     /label=ZKSCAN1 intron 1 partial
     misc_feature    958..997
                     /label=AluSq2
     misc_feature    1027..1044
                     /label=splice site
     misc_feature    1046..1051
                     /label=BsmBI
     misc_feature    1060..1065
                     /label=BsmBI
     misc_feature    1067..1125
                     /label=ZKSCAN1 intron 3 partial
     misc_feature    1067..1072
                     /label=splice site
     misc_feature    1090..1125
                     /label=AluSz
     polyA_signal    1169..1393
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1174..1193)
                     /label=BWS337
     rep_origin      1439..1867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1881..2210
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2061..2196
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2231..2240
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2237..2947
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2492..2501
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    3107..3228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3277..3293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3301..3317
                     /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(3325..3355)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3370..3391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3679..4267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4438..5298)
                     /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(5299..5403)
                     /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 atccactagt ccagtgtggt ggaattcaaa
      961 gtgctgagat tacaggcgtg agccaccacc cccggcccac tttttgtaaa ggtacgtact
     1021 aatgactttt tttttatact tcagggagac ggttgtgtac gtctcggtaa gaagcaagga
     1081 aaagaattag gctcggcacg gtagctcaca cctgtaatcc cagcactcga gtctagaggg
     1141 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1201 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1261 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1321 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1381 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1441 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1501 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1561 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1621 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1681 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1741 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1801 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1861 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1981 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2041 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2101 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2161 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2221 gaccggtgcc gccaccatgg tgagcaaggg cgaggaggat aacatggcct ctctcccagc
     2281 gacacatgag ttacacatct ttggctccat caacggtgtg gactttgaca tggtgggtca
     2341 gggcaccggc aatccaaatg atggttatga ggagttaaac ctgaagtcca ccaagggtga
     2401 cctccagttc tccccctgga ttctggtccc tcatatcggg tatggcttcc atcagtacct
     2461 gccctaccct gacgggatgt cgcctttcca ggccgccatg gtagatggct ccggatacca
     2521 agtccatcgc acaatgcagt ttgaagatgg tgcctccctt actgttaact accgctacac
     2581 ctacgaggga agccacatca aaggagaggc ccaggtgaag gggactggtt tccctgctga
     2641 cggtcctgtg atgaccaact cgctgaccgc tgcggactgg tgcaggtcga agaagactta
     2701 ccccaacgac aaaaccatca tcagtacctt taagtggagt tacaccactg gaaatggcaa
     2761 gcgctaccgg agcactgcgc ggaccaccta cacctttgcc aagccaatgg cggctaacta
     2821 tctgaagaac cagccgatgt acgtgttccg taagacggag ctcaagcact ccaagaccga
     2881 gctcaacttc aaggagtggc aaaaggcctt taccgatgtg atgggcatgg acgagctgta
     2941 caagtaattc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3001 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3061 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3121 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3181 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3241 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3301 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3361 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3481 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3541 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3601 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3661 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3721 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3781 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3841 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3901 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3961 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4021 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4081 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4141 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4201 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4261 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4321 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4381 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4441 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4501 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4561 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4621 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4681 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4741 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4801 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4861 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4921 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4981 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5041 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5101 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5161 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5221 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5281 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5341 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5401 gcgcacattt ccccgaaaag tgccacctga cgtc
//
