LOCUS       Exported                7231 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 7231)
  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..7231
                     /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..3381
                     /codon_start=1
                     /label=Zaphod2
                     /translation="MSEKNFKKLSGSENRKWKAIREENERKLFGSLNKFLEKIDGMPEK
                     KEKTSELLVSAECSTDTSLMEQDVNLPSTSMSTITDEFQEKLEEKKSEESIKPNVLSAD
                     PGTWPCNLNIKQRDTLVENYPPQVRNFNFPKDSTGRKFSETYYTRILPNGEKITRSWLL
                     YSTSKDSVFCLYCKLFGDGKNQLKNENGCKDWQHLSHILSKHEESEMHINNSVKYSKLK
                     SDLKKYITIDAAEQRLYENEEKYWRAVVERIIYMIQFLARQCLAFRGSSKKLYERDNGN
                     FLHLAETIAKFDNIMAEHLCRIQTSTPNNITHYLGDEIQNQIIKIIADAMNEIILNKVR
                     NAKYYSIILDTTPDVSHVEQITIIVRFVDFQNANISICEHFGGFCPVDDITGEGLYTFL
                     VDYLQKLNLDKNNLRGQGYDNATNMHGKHNGLQQKILEKYSRAFYIPCSAHSLNLVVND
                     AAKISFDTTEFFYIVQELYNYFSASTKRWSVLKTHLPNINLKPLSNTRWESRVDAIMPL
                     KTQLEAVCNALLEIVDDINKDIDSRMQAKGILSKISTFKFICSVLIWHDILSKINVISK
                     MMQEPKFNVHNALESIKGIKKFFEEKRSDDVYEKYIIDAKQIAESLDIDANFSNVENTR
                     KRRKKTHFDCESHDQPIYDPKLNFKINFFFSILDTTISSLKERFTILKEYHGLFSVLDD
                     ICNLKNYNERELLQLCKNLQLKLTDEHSKECDIDGTSMYEELKTLSCFLKSSMTPVEVL
                     QYICSNNLPDTFPNTFVALRIYLTLPISVASGERSFSKLKFIKNYLRSTMSQERLSNLA
                     TISIENEILEQVSIETVIKNFISMKARRVTIL"
     promoter        complement(3419..3437)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3463..3687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3733..4161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4175..4504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4355..4490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4670..4791
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4840..4856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4864..4880
                     /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(4888..4918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4933..4954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6001..6861)
                     /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(6862..6966)
                     /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 gcgagaagaa cttcaaaaaa ctgagcggca gcgagaaccg caaatggaaa
      901 gcgattcgcg aagagaacga acgcaaactg tttggcagcc tgaacaaatt cctggaaaag
      961 atcgatggca tgccggaaaa gaaagaaaaa accagcgagc tgctggttag cgcagaatgc
     1021 agcaccgaca cctcactgat ggagcaggat gtgaacttac cgagcaccag catgagcacc
     1081 atcaccgatg aatttcagga aaagctggag gaaaaaaaga gcgaggagag cattaaacct
     1141 aacgtgctga gcgcggatcc tggtacttgg ccttgcaacc tgaacattaa gcagcgcgat
     1201 accctggtgg aaaactatcc tccgcaggtg cgcaacttca actttcctaa agatagcacc
     1261 ggccgcaagt tcagcgaaac ctactatacc cgcatcctgc ctaacggcga aaaaattacc
     1321 cgcagctggc tgttatatag caccagcaag gatagcgtgt tttgcctgta ctgcaaactg
     1381 tttggcgatg gcaaaaacca gctgaaaaac gaaaacggct gcaaagattg gcagcatctg
     1441 agccatatcc tgagcaaaca tgaagagagc gagatgcata tcaacaacag cgtgaaatac
     1501 agcaaactga agagcgacct gaagaagtac atcaccattg atgccgcgga acagcgcctg
     1561 tatgaaaacg aagagaaata ttggcgcgcg gtggtggagc gcattattta catgatccag
     1621 tttctggcgc gccaatgctt agcgtttcgt ggcagcagca aaaagctgta cgagcgcgac
     1681 aacggcaact ttctgcatct ggcggaaacc attgcgaaat tcgataacat tatggcggag
     1741 catctgtgcc gtattcagac cagcaccccg aacaacatta cccattatct gggcgacgag
     1801 attcagaacc agatcatcaa gattattgcc gatgcgatga acgagatcat tctgaacaaa
     1861 gtgcgcaacg cgaaatatta tagcatcatc ctggatacca ccccggatgt gtcacatgtg
     1921 gagcagatca ccattattgt gcgctttgtg gattttcaga acgccaacat tagcatctgc
     1981 gaacattttg gcggcttttg cccggtggat gatattaccg gcgaaggcct gtataccttt
     2041 ctggtggatt acctgcagaa gctgaacctg gataaaaaca acctgcgcgg ccagggttat
     2101 gataacgcca ccaacatgca tggcaaacat aacggcctgc agcagaaaat tctggaaaag
     2161 tatagccgcg ccttctatat tccgtgcagc gcgcatagct taaacctggt ggtgaacgat
     2221 gcggcgaaga ttagcttcga taccaccgag tttttctaca ttgtgcagga actgtacaac
     2281 tattttagcg cgagcaccaa acgttggagc gtgctgaaaa cccacctgcc gaacattaac
     2341 ctgaaacctc tgagcaacac ccgttgggaa agccgcgttg atgcgatcat gccgctgaaa
     2401 acccagttag aagcggtgtg caacgcgctg ctggaaattg tggacgacat caacaaagat
     2461 attgacagcc gcatgcaggc gaaaggcatt ctgagcaaaa tcagcacctt caagttcatt
     2521 tgcagcgtgc tgatttggca tgacattctg agcaagatca acgtgattag caagatgatg
     2581 caggagccta aatttaacgt gcacaacgcc ctggaaagca ttaaaggcat caagaagttc
     2641 tttgaggaga aacgcagcga tgatgtgtat gagaagtata ttatcgacgc gaagcagatt
     2701 gcggaaagcc tggatattga cgccaacttc agcaacgtgg aaaacactcg caagcgccgc
     2761 aagaagaccc attttgattg cgaaagccat gatcagccta tttatgaccc gaagctgaac
     2821 tttaagatca acttcttctt cagcattctg gacaccacca ttagcagcct gaaagaacgc
     2881 tttaccatcc tgaaagagta tcacggcctg tttagcgtgc tggatgatat ttgcaacctg
     2941 aagaactata acgaacgcga actgctgcag ctgtgcaaaa acctgcagct gaaactgacc
     3001 gatgaacata gcaaggaatg cgatattgac ggcaccagca tgtatgagga actgaagacc
     3061 ctgagctgct ttctgaagag cagcatgacc cctgttgagg tgctgcagta catttgcagc
     3121 aacaacctgc cggacacctt tccgaacacc tttgtggccc tgcgcattta tctgaccctg
     3181 ccgatcagcg ttgcaagcgg cgaacgcagc tttagcaagc tgaagttcat caagaactat
     3241 ctgcgcagca ccatgagcca agaacgcctg agcaacctgg ccaccattag cattgaaaac
     3301 gagattctgg agcaggtgag cattgaaacc gtgatcaaaa acttcatcag catgaaggcg
     3361 cgccgtgtga ccatactgtg atgagcggcc gctcgagcat gcatctagag ggccctattc
     3421 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     3481 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3541 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3601 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3661 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3721 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3781 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3841 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3901 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3961 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4021 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4081 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4141 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4201 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4261 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4321 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4381 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4441 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4501 caaaaagctc cccgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4561 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4621 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4681 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4741 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4801 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4861 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4921 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4981 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5041 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7021 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7081 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7141 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     7201 cttcgcgatg tacgggccag atatacgcgt t
//
