LOCUS       CDS-G016_FLAG_SN        6642 bp DNA     circular SYN 09-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6642)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(518..539)
                     /label=ITR1-seq
     primer_bind     complement(532..549)
                     /label=ITR1-seq-R
     promoter        701..978
                     /label=chicken beta-actin promoter
     regulatory      1394..1403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1403..1426
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2252..2305
                     /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"
     misc_feature    3036..3624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3507..3518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3667..3788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4098..4114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4256..4711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4289..4307)
                     /label=ITR2-seq-R
     promoter        4737..4841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4842..5702
                     /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      5873..6461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatcgta ccattgacgt caataatgac gtatgttccc
      481 atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt acggtaaact
      541 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      601 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      661 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      721 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      781 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      841 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      901 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      961 cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc
     1021 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     1081 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     1141 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     1201 cggctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1261 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1321 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1381 aagaattgga tccgccacca tggattacaa ggacgatgac gataagggcg gcgctagcgg
     1441 aggcagcagc gaggtgggcc ctgccctgtt ccaaagcgac atgacattca aaatcttcat
     1501 cgacggcgag gtgaacggcc agaagtttac catcgtggcc gatggctcca gcaagttccc
     1561 ccacggcgac ttcaacgctc acgccgtttg tgaaacaggc aagctgccaa tgagctggaa
     1621 gcctatctgc cacctgatcc agtggggcga gcctttcttc gctagatacc ccgatggaat
     1681 cagccacttc gcccaggagt gcttccccga gggcctgagc atcgatagaa ccgtgcggtt
     1741 tgaaaacgac ggcaccatga catctcacca cacctacgag ctggacgaca cctgcgtggt
     1801 cagccggatc accgtgaatt gcgacggctt tcaacctgac ggccccatta tgcgggacca
     1861 gctggtggac atcctgccta gcgagaccca catgttccct catggcccaa acgccgtgcg
     1921 ccagaccgcc accatcggct tcaccacagc cgacggcgga aaaatgatgg gccacttcga
     1981 cagcaagatg acctttaacg gcagcagagc catcgagatc cctggacctc acttcgtgac
     2041 cattatcaca aagcagacaa gagatacatc cgataaaaga gatcacgtgt gccagcggga
     2101 agtcgcctac gcccacagcg tgccaagaat caccagcgcc atcggcagtg acgaggactc
     2161 tggcggcctt aagggcagcg gcggaggcaa gctgaaccct cctgatgaga gcggccctgg
     2221 ttgtatgagc tgctgcgtgc tgagcaggtc cgaaggcaga ggctctctgc tgacatgtgg
     2281 cgacgtggaa gagaatcccg gacccgctcc tggcagcacc atggtgtcca agggggagga
     2341 agtgatcaag gagttcatga gatttaaggt gcggatggaa ggctctatga acggacatga
     2401 gttcgagatc gaaggcgagg gcgaaggcag accttatgag gggacccaga ccgccaagct
     2461 taaagtgacc aagggcggac ctctgccctt cgcctgggac atcctcagcc ctcaattcat
     2521 gtacggtagc aaggcttacg tcaaacaccc tgctgatata cctgattaca aaaaactgtc
     2581 atttcctgaa ggatttaagt gggaacgagt aatgaatttt gaggatggtg ggctggtcac
     2641 tgtcacgcaa gactcctctc tgcaggacgg caccctgatt tacaaggtga agatgcgcgg
     2701 aacaaatttt ccccccgatg gccccgtgat gcagaagaag accatggggt gggaggcctc
     2761 tacagaacga ctgtaccccc gcgatggcgt tcttaaaggc gaaatccatc aggcgcttaa
     2821 actcaaagat ggcgggcact acctggttga gttcaaaact atatatatgg ccaaaaagcc
     2881 ggttcagctg cctggctact actacgtgga caccaagttg gacatcacga gtcacaacga
     2941 ggactacaca atcgtcgagc agtacgaacg ctcagaaggt cggcatcacc tgttcttgta
     3001 tggcatggat gagctgtata agtaagctta tcgataatca acctctggat tacaaaattt
     3061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3421 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3601 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag
     3661 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3721 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3781 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3841 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     3901 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3961 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4021 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4081 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4141 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4201 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     4261 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4321 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4381 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4441 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4501 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4561 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4621 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4681 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4741 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4801 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4861 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4921 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4981 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5041 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5101 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5161 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5221 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5281 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5341 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5401 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5461 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5521 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5581 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5641 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5701 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5761 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5821 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5881 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5941 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6001 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6061 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6121 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6181 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6241 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6301 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6361 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6421 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6481 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6541 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6601 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
