LOCUS       62056_20490        6335 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6335)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1338..2054
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2055..2081
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    complement(2101..2125)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2178..2219
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2263..2487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2557..2590
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3048..3064)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3538..3642
                     /label=AmpR promoter
     CDS             3643..4500
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4674..5262
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5550..5571
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5586..5616
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5624..5640
                     /label=lac operator
                     /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     5648..5664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6335
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1381 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1441 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1501 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1561 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1621 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1681 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1741 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1801 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1861 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1921 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1981 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2041 acgagctgta caagtaccca tacgacgtcc cagactacgc ttgaaagggt gggcgcgccg
     2101 acccagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     2161 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     2221 gtaccggtta gtaatgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2281 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2341 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2401 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2461 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2521 gggggtatcc ccacgcgccc tgtagcggcg cattaaataa cttcgtataa tgtatgctat
     2581 acgaagttat atcgtatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     2641 tccacaccct aactgacaca cattccacag aattccttga aatacatcca caggtacacc
     2701 tccaattgac tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat
     2761 tttctggaat tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga
     2821 cccactggaa ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt
     2881 ggaaaaatga cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg
     2941 tttgttaaca agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag
     3001 tgtatgtaaa cttccgactt caactgtagg gtaccgagct cgaattcact ggccgtcgtt
     3061 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3121 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3181 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3241 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3301 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3361 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3421 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3481 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3661 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3721 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3781 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3841 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3901 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3961 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4021 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4081 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4141 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4201 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4261 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4321 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4381 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4441 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4501 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4801 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5281 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5341 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5401 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     5461 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5521 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     5581 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     5641 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac
     5701 tctagaccta tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac
     5761 tcgtttttca actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt
     5821 aggacatcta ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt
     5881 atttcactta taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag
     5941 ttgactgtgc ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc
     6001 ttctgataga ctaattgaca tcatttgagt caattggagg tgtacctgtg gatgtatttc
     6061 aaggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     6121 agcggccgct agctcgagat cttaattaaa ctagtatcga tataacttcg tataatgtat
     6181 gctatacgaa gttatatgta cgggccagat atacgcgttg acattgatta ttgactaggc
     6241 ttttgcaaaa agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     6301 aatggacctt ctaggtcttg aaaggagtgg gaatt
//
