LOCUS       Exported                5274 bp ds-DNA     circular SYN 19-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  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..5274
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     133..152
                     /label=BWS533
     promoter        234..734
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     766..785
                     /label=BWS335
     misc_feature    798..884
                     /label=ZKSCAN1 intron 1 partial
     misc_feature    798..837
                     /label=AluSq2
     misc_feature    867..884
                     /label=splice site
     misc_feature    886..891
                     /label=BsmBI
     misc_feature    900..905
                     /label=BsmBI
     misc_feature    907..965
                     /label=ZKSCAN1 intron 3 partial
     misc_feature    907..912
                     /label=splice site
     misc_feature    930..965
                     /label=AluSz
     polyA_signal    1009..1233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1014..1033)
                     /label=BWS337
     rep_origin      1279..1707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1721..2050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1901..2036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2071..2080
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2077..2787
                     /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      2332..2341
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    2947..3068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3117..3133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3141..3157
                     /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(3165..3195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3210..3231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3519..4107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4278..5138)
                     /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(5139..5243)
                     /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 cgtttgcgcc
      241 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
      301 aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc
      361 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
      421 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctta
      481 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
      541 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
      601 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
      661 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
      721 acctctctcc ccaggctagc gtttaaactt aagcttggta ccgagctcgg atccactagt
      781 ccagtgtggt ggaattcaaa gtgctgagat tacaggcgtg agccaccacc cccggcccac
      841 tttttgtaaa ggtacgtact aatgactttt tttttatact tcagggagac ggttgtgtac
      901 gtctcggtaa gaagcaagga aaagaattag gctcggcacg gtagctcaca cctgtaatcc
      961 cagcactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     1021 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1081 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1141 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1201 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1261 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1321 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1381 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1441 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1501 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1561 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1621 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1681 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     1741 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1801 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1861 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     1921 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1981 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2041 cttttgcaaa aagctcccgg gaccggtgcc gccaccatgg tgagcaaggg cgaggaggat
     2101 aacatggcct ctctcccagc gacacatgag ttacacatct ttggctccat caacggtgtg
     2161 gactttgaca tggtgggtca gggcaccggc aatccaaatg atggttatga ggagttaaac
     2221 ctgaagtcca ccaagggtga cctccagttc tccccctgga ttctggtccc tcatatcggg
     2281 tatggcttcc atcagtacct gccctaccct gacgggatgt cgcctttcca ggccgccatg
     2341 gtagatggct ccggatacca agtccatcgc acaatgcagt ttgaagatgg tgcctccctt
     2401 actgttaact accgctacac ctacgaggga agccacatca aaggagaggc ccaggtgaag
     2461 gggactggtt tccctgctga cggtcctgtg atgaccaact cgctgaccgc tgcggactgg
     2521 tgcaggtcga agaagactta ccccaacgac aaaaccatca tcagtacctt taagtggagt
     2581 tacaccactg gaaatggcaa gcgctaccgg agcactgcgc ggaccaccta cacctttgcc
     2641 aagccaatgg cggctaacta tctgaagaac cagccgatgt acgtgttccg taagacggag
     2701 ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaaaggcctt taccgatgtg
     2761 atgggcatgg acgagctgta caagtaattc gaaatgaccg accaagcgac gcccaacctg
     2821 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     2881 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     2941 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3001 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3061 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     3121 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3181 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
