LOCUS       Exported                4985 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 4985)
  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..4985
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     133..152
                     /label=BWS533
     promoter        234..445
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     477..496
                     /label=BWS335
     misc_feature    509..595
                     /label=ZKSCAN1 intron 1 partial
     misc_feature    509..548
                     /label=AluSq2
     misc_feature    578..595
                     /label=splice site
     misc_feature    597..602
                     /label=BsmBI
     misc_feature    611..616
                     /label=BsmBI
     misc_feature    618..676
                     /label=ZKSCAN1 intron 3 partial
     misc_feature    618..623
                     /label=splice site
     misc_feature    641..676
                     /label=AluSz
     polyA_signal    720..944
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(725..744)
                     /label=BWS337
     rep_origin      990..1418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1432..1761
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1612..1747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1782..1791
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1788..2498
                     /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      2043..2052
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    2658..2779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2828..2844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2852..2868
                     /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(2876..2906)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2921..2942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3230..3818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3989..4849)
                     /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(4850..4954)
                     /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 cgtgggcaga
      241 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
      301 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      361 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      421 caacgggttt gccgccagaa cacaggctag cgtttaaact taagcttggt accgagctcg
      481 gatccactag tccagtgtgg tggaattcaa agtgctgaga ttacaggcgt gagccaccac
      541 ccccggccca ctttttgtaa aggtacgtac taatgacttt ttttttatac ttcagggaga
      601 cggttgtgta cgtctcggta agaagcaagg aaaagaatta ggctcggcac ggtagctcac
      661 acctgtaatc ccagcactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
      721 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
      781 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
      841 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
      901 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
      961 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     1021 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1081 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1141 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1201 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1261 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1321 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1381 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     1441 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1501 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1561 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     1621 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     1681 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     1741 ggaggcctag gcttttgcaa aaagctcccg ggaccggtgc cgccaccatg gtgagcaagg
     1801 gcgaggagga taacatggcc tctctcccag cgacacatga gttacacatc tttggctcca
     1861 tcaacggtgt ggactttgac atggtgggtc agggcaccgg caatccaaat gatggttatg
     1921 aggagttaaa cctgaagtcc accaagggtg acctccagtt ctccccctgg attctggtcc
     1981 ctcatatcgg gtatggcttc catcagtacc tgccctaccc tgacgggatg tcgcctttcc
     2041 aggccgccat ggtagatggc tccggatacc aagtccatcg cacaatgcag tttgaagatg
     2101 gtgcctccct tactgttaac taccgctaca cctacgaggg aagccacatc aaaggagagg
     2161 cccaggtgaa ggggactggt ttccctgctg acggtcctgt gatgaccaac tcgctgaccg
     2221 ctgcggactg gtgcaggtcg aagaagactt accccaacga caaaaccatc atcagtacct
     2281 ttaagtggag ttacaccact ggaaatggca agcgctaccg gagcactgcg cggaccacct
     2341 acacctttgc caagccaatg gcggctaact atctgaagaa ccagccgatg tacgtgttcc
     2401 gtaagacgga gctcaagcac tccaagaccg agctcaactt caaggagtgg caaaaggcct
     2461 ttaccgatgt gatgggcatg gacgagctgt acaagtaatt cgaaatgacc gaccaagcga
     2521 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     2581 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     2641 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2701 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2761 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     2821 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2881 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2941 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3001 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3061 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3421 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3661 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4981 acgtc
//
