LOCUS       Exported                6835 bp ds-DNA     circular SYN 03-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 3, 2020 from SnapGene 5.1.5
            https://www.snapgene.com
COMMENT     TTCTAGTTGCCAGCCATCT
FEATURES             Location/Qualifiers
     source          1..6835
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    659..1462
                     /locus_tag="HA-L"
                     /label=HA-L
     misc_feature    1463..1488
                     /locus_tag="Splice/acceptor"
                     /label=Splice/acceptor
     CDS             1512..1565
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1575..2174
                     /codon_start=1
                     /locus_tag="Puro/Resistance Gene"
                     /label=Puro/Resistance Gene
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1717..1737)
                     /label=Puro_Reverse_Primer
     polyA_signal    2328..2439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2529..2740
                     /locus_tag="4X ZF1 BD"
                     /label=4X ZF1 BD
     misc_feature    2568..2587
                     /locus_tag="binding site for ZF1"
                     /label=binding site for synthetic Zing Finger DBD
     misc_feature    2606..2625
                     /label=binding site for synthetic Zing Finger DBD
     misc_feature    2644..2663
                     /label=binding site for synthetic Zing Finger DBD
     misc_feature    2682..2701
                     /label=binding site for synthetic Zing Finger DBD
     misc_feature    2747..2811
                     /locus_tag="pMiniCMV"
                     /label=pMiniCMV
     misc_feature    2827..2832
                     /locus_tag="Kozak"
                     /label=Kozak
     CDS             2833..3549
                     /codon_start=1
                     /locus_tag="GFP"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    3559..3680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3781..4617
                     /locus_tag="HA-R"
                     /label=HA-R
     promoter        complement(4647..4665)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4686..4702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4710..4726
                     /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(4734..4764)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4779..4800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5088..5676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5847..6707)
                     /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(6708..6812)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctg
      661 ctttctctga ccagcattct ctcccctggg cctgtgccgc tttctgtctg cagcttgtgg
      721 cctgggtcac ctctacggct ggcccagatc cttccctgcc gcctccttca ggttccgtct
      781 tcctccactc cctcttcccc ttgctctctg ctgtgttgct gcccaaggat gctctttccg
      841 gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct gagcggatcc tccccgtgtc
      901 tgggtcctct ccgggcatct ctcctccctc acccaacccc atgccgtctt cactcgctgg
      961 gttccctttt ccttctcctt ctggggcctg tgccatctct cgtttcttag gatggccttc
     1021 tccgacggat gtctcccttg cgtcccgcct ccccttcttg taggcctgca tcatcaccgt
     1081 ttttctggac aaccccaaag taccccgtct ccctggcttt agccacctct ccatcctctt
     1141 gctttctttg cctggacacc ccgttctcct gtggattcgg gtcacctctc actcctttca
     1201 tttgggcagc tcccctaccc cccttacctc tctagtctgt gctagctctt ccagccccct
     1261 gtcatggcat cttccagggg tccgagagct cagctagtct tcttcctcca acccgggccc
     1321 ctatgtccac ttcaggacag catgtttgct gcctccaggg atcctgtgtc cccgagctgg
     1381 gaccacctta tattcccagg gccggttaat gtggctctgg ttctgggtac ttttatctgt
     1441 cccctccacc ccacagtggg gcctgacctc ttctcttcct cccacagggc ctcgagagat
     1501 ctggcagcgg agagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     1561 gccctaggct cgaaatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     1621 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     1681 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     1741 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     1801 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     1861 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     1921 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     1981 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     2041 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     2101 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     2161 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     2221 cccatggctc cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag
     2281 gcccaccgac tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2341 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2401 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2461 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2521 caaccggtcg ggtttcgtaa caatcgcatg aggattcgca acgccttcgt cgaagtcgaa
     2581 gtcgacctcc cgtctcagta aaggtcgtcg aagtcgaagt cgaccaatcg gactgccttc
     2641 gtacgtcgaa gtcgaagtcg accgtatcag tcgcctcgga acgtcgaagt cgaagtcgac
     2701 cattcgtaag aggctcactc tcccttacac ggagtggata actagttagg cgtgtacggt
     2761 gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg aacgcgtacc
     2821 ggtgtcgcca ccatgagtaa aggcgaagag ctctttacag gtgtggtgcc catccttgtc
     2881 gaacttgatg gggacgtgaa tggccataaa ttttctgtaa gcggtgaagg agagggcgat
     2941 gcaacttacg gtaagctcac actcaagttt atctgcacca caggtaaact gcccgtgccc
     3001 tggccgaccc tggtcacgac gttcggttac ggtgtgcaat gctttgcgcg gtatccagat
     3061 catatgaaac aacatgactt ttttaagtcc gcgatgccgg aggggtatgt ccaggaaagg
     3121 actatcttct tcaaggatga cggtaactat aagacacggg ccgaggttaa attcgagggt
     3181 gacacccttg tcaaccgcat cgaattgaag gggatagact tcaaggaaga tgggaatata
     3241 ctggggcaca aactggaata taattataat agtcacaacg tttatattat ggcggataag
     3301 caaaagaacg gaatcaaggt gaacttcaaa ataaggcaca acatagaaga cggatccgta
     3361 caattggccg atcactatca acaaaatacg ccaatagggg acggcccagt ccttctcccc
     3421 gacaatcatt atctgtctac tcaaagtgcg ctctcaaaag acccaaacga gaaaagagat
     3481 cacatggttc tgctggaatt tgtgacagcc gctggaatca cacacggaat ggacgaactt
     3541 tacaaataac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3601 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3661 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3721 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt agcggccgct
     3781 actagggaca ggattggtga cagaaaagcc ccatccttag gcctcctcct tcctagtctc
     3841 ctgatattgg gtctaacccc cacctcctgt taggcagatt ccttatctgg tgacacaccc
     3901 ccatttcctg gagccatctc tctccttgcc agaacctcta aggtttgctt acgatggagc
     3961 cagagaggat cctgggaggg agagcttggc agggggtggg agggaagggg gggatgcgtg
     4021 acctgcccgg ttctcagtgg ccaccctgcg ctaccctctc ccagaacctg agctgctctg
     4081 acgcggctgt ctggtgcgtt tcactgatcc tggtgctgca gcttccttac acttcccaag
     4141 aggagaagca gtttggaaaa acaaaatcag aataagttgg tcctgagttc taactttggc
     4201 tcttcacctt tctagtcccc aatttatatt gttcctccgt gcgtcagttt tacctgtgag
     4261 ataaggccag tagccagccc cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt
     4321 ggaaaactcc ctttgtgaga atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct
     4381 actccctttc tctttctcca tccttctttc cttaaagagt ccccagtgct atctgggaca
     4441 tattcctccg cccagagcag ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt
     4501 tgagtccttg gcaagcccag gagaggcgct caggcttccc tgtccccctt cctcgtccac
     4561 catctcatgc ccctggctct cctgcccctt ccctacaggg gttcctggct ctgctctcca
     4621 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     4681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4921 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5521 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5881 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5941 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6001 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6061 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6721 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6781 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
