LOCUS       62056_20495        5438 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 5438)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5438
                     /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"
     polyA_signal    1366..1590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1660..1693
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2151..2167)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2641..2745
                     /label=AmpR promoter
     CDS             2746..3603
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3777..4365
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4653..4674
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4689..4719
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4727..4743
                     /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     4751..4767
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5438
                     /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 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     1381 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1441 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1501 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     1561 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     1621 ctagggggta tccccacgcg ccctgtagcg gcgcattaaa taacttcgta taatgtatgc
     1681 tatacgaagt tatatcgtat gcatgctttg catacttctg cctgctgggg agcctgggga
     1741 ctttccacac cctaactgac acacattcca cagaattcct tgaaatacat ccacaggtac
     1801 acctccaatt gactcaaatg atgtcaatta gtctatcaga agcttctaaa gccatgacat
     1861 cattttctgg aattttccaa gctgtttaaa ggcacagtca acttagtgta tgtaaacttc
     1921 tgacccactg gaattgtgat acagtgaatt ataagtgaaa taatctgtct gtaaacaatt
     1981 gttggaaaaa tgacttgtgt catgcacaaa gtagatgtcc taactgactt gccaaaacta
     2041 ttgtttgtta acaagaaatt tgtggagtag ttgaaaaacg agttttaatg actccaactt
     2101 aagtgtatgt aaacttccga cttcaactgt agggtaccga gctcgaattc actggccgtc
     2161 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2221 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2281 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2341 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2401 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     2581 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2641 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2701 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2761 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2821 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2881 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2941 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3001 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3061 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3121 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3181 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3241 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3301 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3361 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3421 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3481 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3541 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3601 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3661 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3901 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4441 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4501 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4561 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4621 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4681 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4741 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc
     4801 gactctagac ctatacagtt gaagtcggaa gtttacatac acttaagttg gagtcattaa
     4861 aactcgtttt tcaactactc cacaaatttc ttgttaacaa acaatagttt tggcaagtca
     4921 gttaggacat ctactttgtg catgacacaa gtcatttttc caacaattgt ttacagacag
     4981 attatttcac ttataattca ctgtatcaca attccagtgg gtcagaagtt tacatacact
     5041 aagttgactg tgcctttaaa cagcttggaa aattccagaa aatgatgtca tggctttaga
     5101 agcttctgat agactaattg acatcatttg agtcaattgg aggtgtacct gtggatgtat
     5161 ttcaagggga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5221 tgcagcggcc gctagctcga gatcttaatt aaactagtat cgatataact tcgtataatg
     5281 tatgctatac gaagttatat gtacgggcca gatatacgcg ttgacattga ttattgacta
     5341 ggcttttgca aaaagctttg caaagatgga taaagtttta aacagagagg aatctttgca
     5401 gctaatggac cttctaggtc ttgaaaggag tgggaatt
//
