LOCUS       Exported                7333 bp ds-DNA     circular SYN 22-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7333)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7333
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             847..3483
                     /codon_start=1
                     /label=hAT-4_Crp
                     /translation="MSGYKYKSGSQKHKEVKQRELKEAKGHQSILQFFKTDKAFLQSGS
                     GNTLTREVDEQKQDGNDFNIETVSVATDDEEKLPDASCDSNLMLSKDSQVISPLAPLRS
                     EEGLQKTGESSTTLSDSCFFKDPGLWPKLITDSVRQKIVLSGVLTFEEMWKLAKSIPKD
                     VGGHKFPEFLLKCKTSNGEQLPRDWLVWSPTNQALYCFPCWLLSEGRQKQCSMLAKSEG
                     YSPNVKKWQKLYNELPEHEKSIEHKTCYCQWKELQQSLSGGHGVDSEVQKQILTEATRW
                     KEILERLLDVTLYLASRGLAFQGDNNLIGDPHNGNFLGTLELIAHYDSITRDHLAKVKQ
                     KQTEGKTMQGQAHYLSWESQNEFTELCGKSVQKAILAEREKAIYYSIICDATPDVSHQE
                     QHVLIFQFVVHNKDTGTFNIKERFFEFINFDLKNGEQIAEMLIASLKRHNIPLDDCRGQ
                     GFDNGSNMSGKIKGVQACIMSKNELAVYSPCASHSLNLVGVNAAAACTETLTFFGCVQQ
                     LFVFFSASPVRWKIITDKIHCSLHSLSDTCWSARADAIRPLTKHLPGILEALECALELN
                     LTHKAQSQLEALKRYFSSFSCLLLASFWYKVLSSIDDRNKILQARGISLESEAALIADI
                     IEEVKQLRDQWPKILQEACLVAEGMGILPQLPKKRGKKQKRFHDDTAEESCSTISENDP
                     ESLFRRNVFDATLDNVIYGLERRFKATNNICFTFRSVLKYVTLEPEELDKASKCLLHRY
                     PTDLSENFPCEMLHLKKNFKSTFGKEYELPPLILLNEIYKKQLQLIFSEVCIAIRIFCT
                     LPVTVAEAERSFSKLALIKNCRRSTMGQERLSHLAILSIECELAREIDFGEIIHDFAMK
                     KARKIKL"
     promoter        complement(3521..3539)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3565..3789
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3835..4263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4277..4606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4457..4592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4772..4893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4942..4958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4966..4982
                     /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(4990..5020)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5035..5056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5344..5932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6103..6963)
                     /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(6964..7068)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacgtc cgcgcagggc aatagcagag ctctctggct
      721 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      781 ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcgcc
      841 gccaccatga gcggctacaa atataaaagc ggcagccaga agcataaaga ggtgaaacag
      901 cgcgaactga aagaagcgaa aggccatcag agcatcctgc agttttttaa aaccgacaag
      961 gcctttctgc aaagcggcag cggcaatacc ttaacccgcg aagtggatga gcagaaacag
     1021 gacggcaacg attttaacat tgaaacggtg agcgtggcca ccgatgatga agagaaactg
     1081 ccggatgcca gctgtgacag caacctgatg ctgagcaagg atagccaggt gattagcccg
     1141 ctggcgcctt tacgtagcga agaaggcctg cagaaaaccg gtgaaagcag caccaccctg
     1201 agcgatagct gcttttttaa agatcctggc ctgtggccga agctgattac cgatagcgtg
     1261 cgccagaaaa ttgtgctgag cggcgtgtta acctttgagg agatgtggaa actggcgaaa
     1321 agcatcccta aagatgtggg cggccataag tttccggagt ttctgctgaa gtgcaaaacc
     1381 agcaacggcg agcaactgcc tcgtgattgg ttagtttgga gcccgaccaa ccaagcgctg
     1441 tattgctttc cgtgctggtt actgtcagaa ggccgccaaa aacagtgtag catgctggcc
     1501 aaaagcgaag gctatagccc gaacgtgaaa aaatggcaga aactgtacaa cgagctgcct
     1561 gagcatgaga aaagcattga acacaagacc tgctattgcc agtggaagga actgcagcag
     1621 agcctgtcag gtggtcatgg cgtggatagc gaagtgcaga aacagattct gaccgaagcg
     1681 acccgctgga aagaaattct ggagcgcctg ctggatgtga ccctgtattt agcgagccgc
     1741 ggtttagcat tccagggcga taacaacctg attggcgatc cgcataacgg caactttctg
     1801 ggcaccctgg agttaatcgc ccattacgat agcatcaccc gcgaccatct ggccaaagtg
     1861 aaacagaagc agaccgaggg caaaaccatg caaggccagg cccattatct gagctgggaa
     1921 agccagaacg aatttaccga actgtgcggc aaaagcgtgc agaaagccat tctggcggag
     1981 cgcgaaaaag cgatttacta cagcatcatc tgcgatgcca ccccggatgt gtcacatcag
     2041 gaacagcatg tgctgatttt ccagttcgtg gtgcataaca aggacaccgg cacctttaac
     2101 attaaagagc gcttcttcga atttatcaac ttcgacctga aaaacggcga acagatcgcc
     2161 gagatgctga ttgcgagcct gaaacgccac aacattccgc tggatgattg tcgtggccag
     2221 ggctttgata acggcagcaa catgagcggc aagattaaag gcgtgcaggc ctgcattatg
     2281 agcaaaaacg aactggccgt gtatagccct tgtgcgagcc atagcttaaa tctggtgggc
     2341 gtgaatgcgg cagcggcatg tactgaaacc ctgacctttt ttggctgcgt gcagcagctg
     2401 tttgtgtttt tcagcgcgag cccggtgcgc tggaaaatta tcaccgacaa gattcattgc
     2461 agcctgcata gcctgagcga tacctgttgg agcgcacgtg cggatgcaat tcgcccgtta
     2521 accaaacacc tgccgggcat tctggaagca ctggaatgcg cgctggaact gaacctgacc
     2581 cataaggcgc agagccaact ggaagcgctg aaacgctact ttagcagctt tagctgctta
     2641 ctgctggcga gcttttggta taaagtgctg agcagcattg atgaccgcaa caaaatttta
     2701 caggcgcgcg gcattagctt agaaagcgag gcagcgctga ttgcggatat tattgaggaa
     2761 gtgaagcagc tgcgtgacca gtggcctaag attctgcagg aggcctgctt agttgcagaa
     2821 ggcatgggca tactgcctca gctgccgaaa aagcgcggca agaaacagaa acgctttcac
     2881 gatgacaccg ccgaagaaag ctgcagcacc attagcgaaa acgacccgga aagcctgttt
     2941 cgccgcaatg tgttcgatgc caccctggac aacgtgatct atggtctgga gcgccgcttt
     3001 aaagcgacca acaacatctg ctttaccttt cgcagcgtgc tgaaatatgt gaccctggag
     3061 cctgaggaac tggataaggc gagcaaatgc ctgctgcatc gctatcctac cgacctgagc
     3121 gaaaactttc cgtgcgaaat gctgcatctg aagaagaact tcaaaagcac cttcggcaaa
     3181 gaatatgaac tgccgccgct gatcttactg aacgagatct acaaaaagca gctgcagctg
     3241 atctttagcg aagtgtgcat cgcgattcgc attttttgca ccctgcctgt gaccgttgca
     3301 gaagcggaac gcagctttag caagctggcg ctgattaaaa actgccgccg cagcactatg
     3361 ggtcaagagc gcttaagcca tctggcgatc ctgagcatcg agtgtgaact ggcgcgcgag
     3421 attgactttg gcgaaatcat ccacgacttc gcgatgaaaa aagcccgcaa gatcaaactg
     3481 tgatgagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     3541 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3601 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3661 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3721 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3781 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3901 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4021 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4141 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4201 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4261 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4321 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4381 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4441 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4501 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4561 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgaaat
     4621 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4681 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4741 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4801 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4861 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4921 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4981 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5041 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5101 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7081 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     7141 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7201 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7261 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     7321 agatatacgc gtt
//
