LOCUS       Exported                6505 bp ds-DNA     circular SYN 26-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6505)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 1, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6505
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        115..614
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             654..1253
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     654..675
                     /label=PuroR FWD(2)
     primer_bind     complement(654..673)
                     /label=EF1a REV
     primer_bind     complement(1237..1255)
                     /label=PuroR REV(2)
     primer_bind     1809..1825
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1845..2852
                     /label=ArtificialTelomere
     primer_bind     complement(2866..2882)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2875..2911
                     /label=PuroR REV(2)
     CDS             complement(2895..3494)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3473..3514)
                     /label=PuroR FWD(2)
     primer_bind     3475..3515
                     /label=EF1a REV
     promoter        complement(3495..3706)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     complement(3689..3732)
                     /label=EF1a FWD
     promoter        complement(3726..3744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3754..3770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3912..4367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4393..4497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4498..5358
                     /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"
     rep_origin      5529..6117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6405..6426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6441..6471
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6479..6495
                     /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)."
     primer_bind     join(6503..6505,1..14)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
       61 agctggagct ccaccgcggt ggcggccgct attcgtcgac ctcgaaattc taccgggtag
      121 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
      181 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
      241 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
      301 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
      361 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
      421 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
      481 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
      541 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
      601 ctccgggcct ttcgacctgc atccatctag atctcgagca gctgaagctt accatgaccg
      661 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
      721 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
      781 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
      841 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
      901 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
      961 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     1021 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     1081 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     1141 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     1201 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     1261 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg atgatatcag
     1321 atccccggga tgcagaaatt gatgatctat taaacaataa agatgtccac taaaatggaa
     1381 gtttttcctg tcatactttg ttaagaaggg tgagaacaga gtacctacat tttgaatgga
     1441 aggattggag ctacgggggt gggggtgggg tgggattaga taaatgcctg ctctttactg
     1501 aaggctcttt actattgctt tatgataatg tttcatagtt ggatatcata atttaaacaa
     1561 gcaaaaccaa attaagggcc agctcattcc tcccactcat gatctataga tctatagatc
     1621 tctcgtggga tcattgtttt tctcttgatt cccactttgt ggttctaagt actgtggttt
     1681 ccaaatgtgt cagtttcata gcctgaagaa cgagatcagc agcctctgtt ccacatacac
     1741 ttcattctca gtattgtttt gccaagttct aattccatca gaagctggtc gactctagat
     1801 gcggccgctc tagaactagt ggatccgtat accatgaatt cgatccctaa ccctaaccct
     1861 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     1921 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     1981 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2041 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2101 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2161 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2221 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2281 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2341 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2401 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2461 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2521 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2581 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2641 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2701 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2761 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2821 aaccctaacc ctaaccctaa ccctaaccct aaccatcaag cttatcgata ccgtcgacct
     2881 cgaggggggg cccgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     2941 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     3001 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     3061 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     3121 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     3181 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     3241 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     3301 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     3361 cgctcgatgt ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
     3421 gcggcgaggg tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc
     3481 ttgtactcgg tcatctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg
     3541 actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg
     3601 acatcacttt cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc
     3661 cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccggta ccgtacccaa
     3721 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     3781 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3841 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3901 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5641 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6181 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6241 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6301 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6361 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6421 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6481 gtgagcggat aacaatttca cacag
//
